24. Rajapakse, S; Qu, D; Ahmed, AS; Rickers-Haunerland, J; Haunerland, NH. (2019) Effects of FABP knockdown on flight performance of the desert locust, Schistocerca gregaria.J. Exp. Biol. 222 Effects of FABP knockdown on flight performance of the desert locust, Schistocerca gregaria
RNAi; Fatty acid binding protein; Insect flight; Lipid transport
During migratory flight, desert locusts rely on fatty acids as their predominant source of energy. Lipids mobilized in the fat body are transported to the flight muscles and enter the muscle cells as free fatty acids. It has been postulated that muscle fatty acid binding protein (FABP) is needed for the efficient translocation of fatty acids through the aqueous cytosol towards mitochondrial beta-oxidation. To assess whether FABP is required for this process, dsRNA was injected into freshly emerged adult males to knock down the expression of FABP. Three weeks after injection, FABP and its mRNA were undetectable in flight muscle, indicating efficient silencing of FABP expression. At rest, control and treated animals exhibited no morphological or behavioral differences. In tethered flight experiments, both control and treated insects were able to fly continually in the initial, carbohydrate-fueled phase of flight, and in both groups, lipids were mobilized and released into the hemolymph. Flight periods exceeding 30 min, however, when fatty acids become the main energy source, were rarely possible for FABP-depleted animals, while control insects continued to fly for more than 2 h. These results demonstrate that FABP is an essential element of skeletal muscle energy metabolism in vivo. DOI PubMed
22. Han, D; Haunerland, NH; Williams, TD. (2009) Variation in yolk precursor receptor mRNA expression is a key determinant of reproductive phenotype in the zebra finch (Taeniopygia guttata).Journal of Experimental Biology 212: 1277-1283 Variation in yolk precursor receptor mRNA expression is a key determinant of reproductive phenotype in the zebra finch (Taeniopygia guttata)
LOW-DENSITY-LIPOPROTEIN; CHICKEN OOCYTE GROWTH; TISSUE-SPECIFIC EXPRESSION; STARLING STURNUS-VULGARIS; GENE FAMILY-MEMBER; VITELLOGENIN RECEPTOR; INDIVIDUAL VARIATION; INTERINDIVIDUAL VARIATION; ONCORHYNCHUS-MYKISS; OVARIAN-FOLLICLES
The vitellogenin/very low density lipoprotein receptor (VTG/VLDL-R), a 95 kDa protein that belongs to the low density lipoprotein receptor gene family, mediates the uptake of yolk precursors by developing follicles during oocyte growth. However, the extent to which variation in VTG/VLDL-R expression plays a role in determining inter-individual variation in reproductive phenotype ( e. g. follicle or egg size) is not known. Here we show that the mRNA sequence of the zebra finch (Taeniopygia guttata) VTG/VLDL-R shows a high degree of sequence identity (92%) with chicken VTG/VLDL-R mRNA. Using quantitative real-time PCR we measured transcriptional expression of VTG/VLDL-R mRNA in various tissues, and for different stages of oocyte growth, in individual female zebra finches. VTG/VLDL-R mRNA was expressed at high levels in vitellogenic oocytes and in skeletal muscle, and was also detectable in liver, but these tissues expressed different splice variants: the short-form LR8-in oocytes and liver, and the LR8+ form in skeletal muscle. There was significant temporal variation in VTG/VLDL-R expression during follicle growth, with highest levels in ovary and a gradual decrease from pre-F3 to F1 vitellogenic follicles. Variation in ovary mRNA expression was correlated with inter-individual variation in clutch size and laying interval. Furthermore, variation in F3 follicle VTG/VLDL-R mRNA expression was correlated with inter-individual variation in egg mass and F1 follicle mass, suggesting that VTG/VLDL receptor mRNA expression is a key determinant of inter-individual variation in reproductive phenotype. DOI
20.Haunerland, NH; Dennis, EA. (2008) The scientific work of Fritz Spener.European Journal of Lipid Science and Technology 110: 3-4 The scientific work of Fritz Spener
19. Qu, H; Cui, L; Rickers-Haunerland, J; Haunerland, NH. (2007) Fatty acid-dependent expression of the muscle FABP gene - comparative analysis of gene control in functionally related, but evolutionary distant animal systems.Molecular and Cellular Biochemistry 299: 45-53 Fatty acid-dependent expression of the muscle FABP gene - comparative analysis of gene control in functionally related, but evolutionary distant animal systems
fatty acid response element; FAAR; fatty acid activated receptor; intracellular lipid binding proteins; fatty acid transport; adaptation; molecular physiology
The heart is the most fatty acid-dependent muscle in mammals, but flight muscles of birds and insects encounter even higher rates of fatty acid oxidation. The amount of the muscle fatty acid binding protein (H-FABP) found in these muscle reflects their metabolic activities, and increased fatty acid metabolism in endurance exercise increases FABP expression further. We have studied the mechanism of fatty acid-dependent expression of the H-FABP gene, taking advantage of the comparative analysis of gene control in functionally related, but evolutionary distant animal systems, i.e., rat heart and locust flight muscle. Luciferase reporter genes with a full-length promoter (similar to 1 kb) from either the locust or the rat were strongly expressed in L6 myoblasts, and the expression of both constructs was markedly increased by fatty acid treatment. Because of its stronger induction by fatty acids and the absence of other vertebrate transcription factor binding sites, the locust promoter was advantageous for the identification of a fatty acid response element (FARE), an inverted repeat of a hexanucleotide half site reminiscent of steroid hormone receptor binding sites (IR-3). All mammalian H-FABP promoters contain similar sequences, however in reverse orientation (everted repeats, ER-3). Deletion of the FARE eliminated the fatty acid inducibility completely for the locust promoter, but only partly for its mammalian analogue, perhaps because of additional factors or more complex interactions. In gel shift studies, the element binds nuclear proteins from both rat cells and locust flight muscle, further attesting to the far-reaching conservation of this mechanism. Two individual proteins bind to the element, with full binding requiring the presence of free fatty acid. Antibodies to PPARs failed to induce a supershift of the protein-DNA complex, indicating that other transcription factors are responsible for the fatty acid-mediated induction of gene expression of H-FABP. DOI
18. Sum, H; Haunerland, NH. (2007) VHDL, a larval storage protein from the corn earworm, Helicoverpa zea, is a member of the vitellogenin gene family.Insect Biochemistry and Molecular Biology 37: 1086-1093 VHDL, a larval storage protein from the corn earworm, Helicoverpa zea, is a member of the vitellogenin gene family
very high-density lipoprotein; storage protein; metamorphosis; vitellogenin; noctuidae
The hemolymph of last instar larvae of the corn earworm, Helicoverpa zea contains a blue very high-density lipoprotein (VHDL) that is selectively taken up into fat body prior to pupation. Its amino-terminal sequence was determined by Edman degradation, and used to design a degenerate primer for PCR amplification. With 5' and 3' RACE techniques, the entire cDNA coding for VHDL was amplified and sequenced. Conceptual translation reveals a 173 kDa protein that contains a 15 amino acid signal sequence immediately before the experimentally determined N-terminus of the mature protein. The protein contains a typical lipoprotein N-terminal domain, and shows high sequence similarity to vitellogenins from Lepidoptera and other insect species. VHDL mRNA was not detectable in adult H. zea, and antibodies raised against VHDL did not react with adult hemolymph or yolk proteins. Therefore VHDL, although a member of the vitellogenin gene family, seems to be distinct from the vitellogenin expressed in adult females. (C) 2007 Elsevier Ltd. All rights reserved. DOI
17. Cheng, CL; Flamarique, IN; Harosi, FI; Rickers-Haunerland, J; Haunerland, NH. (2006) Photoreceptor layer of salmonid fishes: Transformation and loss of single cones in juvenile fish.J Comp Neurol 495: 213-235 Photoreceptor layer of salmonid fishes: Transformation and loss of single cones in juvenile fish
UV cone; blue cone; opsin; apoptosis; cone mosaic; retinal development
The retinas of many vertebrates have cone photoreceptors that express multiple visual pigments. In many of these animals, including humans, the original cones to appear in the retina (which express UV or blue opsin) may change opsin types, giving rise to new spectral phenotypes. Here we used microspectrophotometry and in situ hybridization with cDNA probes to study the distribution of UV and blue cones in the retinas of four species of Pacific salmon (coho, chum, chinook, and pink salmon), in the Atlantic salmon, and in the rainbow/ steelhead trout. In Pacific salmon and in the trout, all single cones express a UV opsin at hatching lambda(max) of the visual pigment similar to 365 nm), and these cones later transform into blue cones by opsin changeover (lambda(max) of the blue visual pigment similar to 434 nm). Cones undergoing UV opsin downregulation exhibit either of two spectral absorbance profiles. The first is characterized by UV and blue absorbance peaks, with blue absorbance dominating the base of the outer segment. The second shows U-V absorbance diminishing from the outer segment tip to the base, with no sign of blue absorbance. The first absorbance profile indicates a transformation from U-V to blue phenotype by opsin changeover, while the second type suggests that the cone is undergoing apoptosis. These two events (transformation and loss of corner cones) are closely associated in time and progress from ventral to dorsal retina. Each double cone member contains green (lambda(max) similar to 510 nm) or red (lambda(max) similar to 565 nm) visual pigment (double cones are green/red pairs), and, like the rods (lambda(max) similar to 508 nm), do not exhibit opsin changeover. Unlike Pacific salmonids, the Atlantic salmon shows a mixture of ILTV and blue cones and a partial loss of corner cones at hatching. This study establishes the UV-to-blue cone transformation as a general feature of retinal growth in Pacific salmonids (genus Oncorhynchus).
16.Haunerland, NH; Spener, F. (2004) Fatty acid-binding proteins - insights from genetic manipulations.Progress in Lipid Research 43: 328-349 Fatty acid-binding proteins - insights from genetic manipulations
Fatty acid-binding proteins (FABPs) belong to the conserved multigene family of the intracellular lipid-binding proteins (iLBPs). These proteins are ubiquitously expressed in vertebrate tissues, with distinct expression patterns for the individual FABPs. Various functions have been proposed for these proteins, including the promotion of cellular uptake and transport of fatty acids, the targeting of fatty acids to specific metabolic pathways, and the participation in the regulation of gene expression and cell growth. Novel genetic tools that have become available in recent years, such as transgenic cell lines, animals, and knock-out mice, have provided the opportunity to test these concepts in physiological settings. Such studies have helped to define essential cellular functions of individual FABP-types or of combinations of several different FABPs. The deletion of particular FABP genes, however, has not led to gross phenotypical changes, most likely because of compensatory overexpression of other members of the iLBP gene family, or even of unrelated fatty acid transport proteins. This review summarizes the properties of the various FABPs expressed in mammalian tissues, and discusses the transgenic and ablation studies carried out to date in a functional context. (C) 2004 Elsevier Ltd. All rights reserved.
15. Persaud, DR; Haunerland, NH. (2004) Cloning and expression of the VHDL receptor from fat body of the corn ear worm, Helicoverpa zea.Journal of Insect Science 4:6. Epub 2004 Feb 27. Cloning and expression of the VHDL receptor from fat body of the corn ear worm, Helicoverpa zea
storage protein receptor; receptor mediated endocytosis; arylphorin; very high density lipoprotein; basic juvenile hormone; suppressible protein; hexamerin; insect metamorphosis
In Noctuids, storage proteins are taken up into fat body by receptor-mediated endocytosis. These include arylphorin and a second, structurally unrelated very high-density lipoprotein (VHDL). Previously, we have isolated a single storage protein receptor from the corn earworm, Helicoverpa zea, which binds both VHDL and arylphorin. The receptor protein is a basic, N-terminally blocked, similar to 80 kDa protein that is associated with fat body membranes. Microsequencing of proteolytic fragments of the isolated receptor protein revealed internal sequences that were used to clone the complete cDNA of the VHDL receptor by 3' and 5' RACE techniques. The receptor protein, when expressed in vitro via a suitable insect expression vector, reacted with antibodies against the native VHDL receptor and bound strongly to its ligand VHDL, thus confirming that the cloned cDNA represents indeed the previously purified VHDL receptor. The receptor protein and a second, similar protein also found associated with the fat body membrane show considerable homology to putative basic juvenile hormone suppressible proteins cloned previously from other Noctuid species. Sequence analysis revealed that the receptor is likely a peripheral membrane protein that may mediate the selective uptake of VHDL.
14. Persaud, DR; Yousefi, V; Haunerland, N. (2003) Efficient isolation, purification, and characterization of the Helicoverpa zea VHDL receptor.Protein Expression and Purification 32: 260-264 Efficient isolation, purification, and characterization of the Helicoverpa zea VHDL receptor
The study of fat body receptors (e.g., VHDL receptor) in Lepidoptera has been irksome due to the fact that isolation and purification of these proteins are difficult and resulted in extremely low yields. A rapid and efficient method is presented for the purification of Helicoverpa zea VHDL receptor by the use of VHDL biotin ligand complexed to streptavidin coated magnetic beads. The technique can be easily applied to other ligands and allows for the purification of membrane proteins with higher yields compared to previously used methods involving immunopurification. Although the purified protein can be characterized by Western and non-radioactive ligand blots using enhanced chemiluminescence (ECL), a non-radioactive ligand blot method using VHDL-FITC is presented, which allows for the quick analysis of the receptor directly from the blot under standard UV light. Sufficient receptor protein has been derived for amino acid analysis, receptor-ligand and xenobiotic binding studies. (C) 2003 Elsevier Inc. All rights reserved.
13. Guglielmo, CG; Haunerland, NH; Hochachka, PW; Williams, TD. (2002) Seasonal dynamics of flight muscle fatty acid binding protein and catabolic enzymes in a migratory shorebird.American Journal of Physiology-Regulatory Integrative and Comparative Physiology 282: R1405-R1413 Seasonal dynamics of flight muscle fatty acid binding protein and catabolic enzymes in a migratory shorebird
endurance exercise; fuel selection; lipid transport; metabolism
We developed an ELISA to measure heart-type fatty acid binding protein (H-FABP) in muscles of the western sandpiper (Calidris mauri), a long-distance migrant shorebird. H-FABP accounted for almost 11% of cytosolic protein in the heart. Pectoralis H-FABP levels were highest during migration (10%) and declined to 6% in tropically wintering female sandpipers. Premigratory birds increased body fat, but not pectoralis H-FABP, indicating that endurance flight training may be required to stimulate H-FABP expression. Juveniles making their first migration had lower pectoralis H-FABP than adults, further supporting a role for flight training. Aerobic capacity, measured by citrate synthase activity, and fatty acid oxidation capacity, measured by 3-hydroxyacyl-CoA- dehydrogenase and carnitine palmitoyl transferase activities, did not change during premigration but increased during migration by 6, 12, and 13%, respectively. The greater relative induction of H-FABP (+70%) with migration than of catabolic enzymes suggests that elevated H-FABP is related to the enhancement of uptake of fatty acids from the circulation. Citrate synthase, 3-hydroxyacyl-CoA-dehydrogenase, and carnitine palmitoyl transferase were positively correlated within individuals, suggesting coexpression, but enzyme activities were unrelated to H-FABP levels.
12. Wu, QW; Chang, WH; Rickers-Haunerland, J; Higo, T; Haunerland, NH. (2002) Characterization of a new fatty acid response element that controls the expression of the locust muscle FABP gene.Molecular and Cellular Biochemistry 239: 173-180 Characterization of a new fatty acid response element that controls the expression of the locust muscle FABP gene
fatty acid binding protein; fatty acid response element; FARE; Schistocerca gregaria; inverted repeat; everted repeat; gene regulation; gel shift
In vertebrate and invertebrate muscles, the expression of fatty acid binding proteins ( FABP) is induced by long chain fatty acids. To identify the fatty acid response elements that mediate this up-regulation, the gene of the FABP expressed in locust flight muscle was cloned, and its upstream sequences analyzed for potential regulatory elements. Comparison with other muscle FABP promoters revealed the presence of a 19-bp imperfect inverted repeat sequence that contains two hexanucleotide half sites (AGTGGT and ATGGGA), interspersed by 3 nucleotides. The promoter activity was studied with reporter gene constructs in L6 myoblasts, in which H-FABP expression is stimulated by long-chain fatty acids in a similar manner as in adult cardiomyocytes. The 19 bp element, located 180 bp upstream of the transcription start site, was found to be essential for the fatty acid induction of gene expression, and gel shift analysis confirmed that this fatty acid response element is capable of binding nuclear proteins both from rat myoblasts and locust muscle in the presence of fatty acids. A similar, but reverse sequence that is present upstream of all mammalian H-FABP promoters may modulate the expression of the rat H-FABP gene.
11. Chang, W; Rickers-Haunerland, J; Haunerland, NH. (2001) Induction of cardiac FABP gene expression by long chain fatty acids in cultured rat muscle cells.Molecular and Cellular Biochemistry 221: 127-132 Induction of cardiac FABP gene expression by long chain fatty acids in cultured rat muscle cells
cardiac FABP; myoblast; myotube; cardiomyocyte; cell culture; fatty acid; gene regulation
The induction of cardiac FABP expression by long-chain fatty acids was measured in cultured rat myoblasts, myotubes and adult cardiomyocytes. With quantitative RT-PCR techniques, the primary transcription product of the FABP gene and the mature mRNA were measured. Incubations of 30 min resulted in a larger than 2-fold increase of the primary transcript in all cells, and FABP mRNA more than doubled in myoblasts and cardiomyocytes after 10 h of fatty acid exposure. The results demonstrate that long chain fatty acids induce the expression of the cardiac FABP gene in muscle cells and their undifferentiated precursors at the level of transcription initiation, suggesting that all factors involved in fatty acid dependent gene induction are already present in myoblasts. Thus, myoblast cell lines should be useful for the characterization of fatty acid response elements that control the expression of the FABP gene.
10. Wu, QW; Andolfatto, P; Haunerland, NH. (2001) Cloning and sequence of the gene encoding the muscle fatty acid binding protein from the desert locust, Schistocerca gregaria.Insect Biochemistry and Molecular Biology 31: 553-562 Cloning and sequence of the gene encoding the muscle fatty acid binding protein from the desert locust, Schistocerca gregaria
FABP; muscle; locust; Drosophila melanogaster; Schistocerca gregaria; regulatory elements; repetitive elements; microsatellite
Muscle fatty acid binding protein (FABP) is a major cytosolic protein in flight muscle of the desert locust. Schistocerca gregaria. FABP expression varies greatly during development and periods of increased fatty acid utilization, but the molecular mechanisms that regulate its expression are not known. In this study, the gene coding for locust muscle FABP was amplified by PCR and cloned, together with 1.2 kb of upstream sequence. The sequence coding for the 607 bp cDNA is interrupted by two introns of 12.7 and 2.9 kb, inserted in analogous positions as the first and third intron of the mammalian homologues. Both introns contain repetitive sequences also found in other locust genes, and the second intron contains a GT-microsatellite, The promoter sequence includes a canonical TATA box 24 bp upstream of the transcription start site. The upstream sequence contains various potential myocyte enhancer sequences and a 160 bp segment that is repeated three times. In database searches in the genome database of Drosophila melanogaster. a gene with the same gene organization and promoter structure was identified, likely the dipteran homologue of muscle FABP. Upstream of both insect genes, a conserved 19 bp inverted repeat sequence was detected. A similar but reverse palindrome is also present upstream of all mammalian heart FABP genes. possibly representing a novel element involved in muscle FABP expression. (C) 2001 Elsevier Science Ltd. All rights reserved.
9. Wu, QW; Haunerland, NH. (2001) A novel fatty acid response element controls the expression of the flight muscle FABP gene of the desert locust, Schistocerca gregaria.European Journal of Biochemistry 268: 5894-5900 A novel fatty acid response element controls the expression of the flight muscle FABP gene of the desert locust, Schistocerca gregaria
fatty acid binding protein; gene regulation; FARE; myoblast; gel-shift analysis
In many tissues, fatty acid binding protein (FABP) expression is stimulated by exposure to elevated fatty acid levels. In contrast to the FABP genes expressed in other tissues, the molecular mechanisms that mediate the upregulation of the muscle FABP gene have not been elucidated. We have studied the expression of locust flight muscle FABP, a protein that is highly homologous to the mammalian H-FABPs. A 130-bp promoter fragment of the locust gene, which includes a canonical TATA box and several GC boxes, is sufficient for the transcription of a reporter gene in mammalian L6 myoblasts. Twofold higher expression rates are observed when the promoter contains 280 bp or more of upstream sequence. Treatment of myoblasts with various fatty acids leads to a marked increase of expression in the longer constructs, but not in the minimal promoter. We have identified a 19-bp inverted repeat (-162/-180) as the element responsible for the fatty acid-mediated induction of gene expression. Deletion of this element eliminates the fatty acid response, and gel shift analysis demonstrates specific binding to nuclear proteins from both L6 myoblasts and locust flight muscle cells. This fatty acid response element bears no similarity to any known transcription factor binding site. A similar palindrome was also found in the promoter of the Drosophila melanogaster muscle FABP gene, and in reverse orientation upstream of all mammalian heart FABP genes. Given the structural and functional conservation of muscle FABPs and their genes, it is possible that this fatty acid response element also modulates the expression of the mammalian H-FABP genes.
8. Lin, HR; Winston, ML; Haunerland, NH; Slessor, KN. (1999) Influence of age and population size on ovarian development, and of trophallaxis on ovarian development and vitellogenin titres of queenless worker honey bee (Hymenoptera : Apidae).Can. Entomol. 131: 695-706 Influence of age and population size on ovarian development, and of trophallaxis on ovarian development and vitellogenin titres of queenless worker honey bee (Hymenoptera : Apidae)
We examined the factors that might influence ovary development in worker honey bees, Apis mellifera L. Queenless workers at different ages (less than or equal to 12 h, and 4, 8, and 21 d) were tested in cages for ovarian development. Newly emerged, 4- and 8-d-old, and 21-d-old workers had medium-, large-, and small-sized ovaries, respectively, suggesting that of the worker ages tested only 4- and 8-d-old workers are likely to become egg layers in a queenless colony. Also, we compared ovarian development of newly emerged workers that were caged for 14 d and allowed to consume either pollen or royal jelly to that of another group of workers similarly caged but screened so that they could only obtain food via trophallaxis from young bees. Ovaries of newly emerged workers that received food from young bees were as well developed as those of newly emerged workers allowed to take pollen or royal jelly directly. Screened workers also had lower but still elevated vitellogenin levels compared with bees having direct access to food. These results indicate that nurse-age bees functioning as pollen-digesting units affect the ovarian development of other workers and to a lesser extent vitellogenesis via food exchange. We compared the influence of group sizes of 25, 125, and 600 bees per cage on ovarian development for 14 d. The two groups of 25 and 125 bees had similar mean ovary scores, and higher scores than a group of 600 bees. Our findings suggest that nurse-age bees could play an important role in mediating worker fertility via trophallaxis, possibly by differentiating worker dominance status, and generally only young workers become fertile when a queen is lost in a colony. Vitellogenin is a more sensitive parameter to measure bee fertility, and might be a useful tool to further explore ovary development and egg laying in worker social insects. We recommend measuring haemolymph vitellogenin titres and (or) oocyte length of workers in a group of 25 bees per cage, supplied with 50% royal jelly in honey as a standard method to assess honey bee worker fertility in future experiments. DOI
7. Zhang, J; Rickers-Haunerland, J; Dawe, I; Haunerland, NH. (1999) Structure and chromosomal location of the rat gene encoding the heart fatty acid-binding protein.European Journal of Biochemistry 266: 347-351 Structure and chromosomal location of the rat gene encoding the heart fatty acid-binding protein
FABP; FISH mapping; heart; MDGI; RT-PCR
The gene coding for rat heart fatty acid-binding protein (FABP), along with 1.2 kb of its 5 '-untranscribed region, was amplified by PCR, cloned and sequenced. As in other FABP genes, the coding sequence is interrupted by three introns of 3.4, 1.4 and 1.1 kb, respectively. Fluorescence ii? situ hybridization mapping revealed that the gene is located on chromosome 5q36. Using intron-specific primers flanking exon 2, unspliced primary transcript RNA:of the FABP gene was detected in a preparation of total RNA isolated from rat heart, proving that the cloned gene is expressed in adult cardiac tissue.
6. Ando, S; Xu, XH; Tibbits, GF; Haunerland, NH. (1998) Cloning and sequencing of complementary DNA for fatty acid binding protein from rainbow trout heart.Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology 119: 213-217 Cloning and sequencing of complementary DNA for fatty acid binding protein from rainbow trout heart
fatty acid transport; FABP; fish heart; muscle; Oncorhynchus mykiss; evolution
A cDNA encoding a rainbow trout homologue of mammalian heart fatty acid binding protein (H-FABP) was isolated. The deduced protein sequence is 75% identical to that of rat H-FABP. The structural conservation of H-FABPs and their evolutionary relationship are discussed. (C) 1998 Elsevier Science Inc.
5. Ando, S; Xu, XH; Tibbits, GF; Haunerland, NH. (1998) Cloning and sequencing of complementary DNA for fatty acid binding protein from rainbow trout heart.Comp. Biochem. Physiol. B-Biochem. Mol. Biol. 119: 213-217 Cloning and sequencing of complementary DNA for fatty acid binding protein from rainbow trout heart
fatty acid transport; FABP; fish heart; muscle; Oncorhynchus mykiss; evolution
A cDNA encoding a rainbow trout homologue of mammalian heart fatty acid binding protein (H-FABP) was isolated. The deduced protein sequence is 75% identical to that of rat H-FABP. The structural conservation of H-FABPs and their evolutionary relationship are discussed. (C) 1998 Elsevier Science Inc. DOI PubMed
4. Guglielmo, CG; Haunerland, NH; Williams, TD. (1998) Fatty acid binding protein, a major protein in the flight muscle of migrating Western Sandpipers.Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology 119: 549-555 Fatty acid binding protein, a major protein in the flight muscle of migrating Western Sandpipers
bird; endurance exercise; FABP; flight; heart; lipid transport; migration; muscle
Migratory flight in birds is fueled primarily by fatty acid oxidation imposing a requirement for high rates of fatty acid: (a) transport; (b) uptake; and (c) delivery to intracellular sites of beta-oxidation. Muscle fatty acid binding protein (M-FABP) is a cytosolic protein involved in the intracellular transport of fatty acids. Its expression appears to be correlated with muscle fatty acid oxidation capacity. The M-FABP was isolated for the first time from a long distance migrant bird using: (i) size exclusion; (ii) anion exchange; and (iii) hydroxyapatite chromatography. M-FABP has a molecular weight of approximately 14000 Da and an isoelectric point of pH 4.8. A partial amino acid sequence of the protein demonstrated homology to M-FABPs from other species (80% identical to human heart FABP). It was estimated that M-FABP comprises approximately 14 and 21% of total cytosolic protein of the pectoralis and heart, respectively; the highest values yet reported from any vertebrate muscle. The abundance of M-FABP in these tissues suggests that the protein may play a key role in fatty acid supply during endurance flight. Thus, it is proposed that a seasonal increase in M-FABP expression could be a component of physiological preparation for migration. (C) 1998 Elsevier Science Inc. All rights reserved.
3. Zhang, J; Haunerland, NH. (1998) Transcriptional regulation of FABP expression in flight muscle of the desert locust, Schistocerca gregaria.Insect Biochemistry and Molecular Biology 28: 683-691 Transcriptional regulation of FABP expression in flight muscle of the desert locust, Schistocerca gregaria
fatty acid binding protein; quantitative PCR; RT-PCR; computer simulation; gene expression; nuclear RNA
FABP is the most abundant cytosolic protein in developed fight muscle of adult locusts, but it is completely absent in nymphs. During the two weeks following adult ecdysis, FABP rises to 18% of the total soluble proteins. its mRNA increases steadily up to day 8, before it gradually declines until reaching a low concentration at day 15, which remains constant for the rest of the animal's life. Using a PCR primer combination specific for a 597 bp sequence of intron 1, we have developed a reverse transcription PCR assay to quantify the amount of primary transcript present in muscle tissue at various ages. The FABP gene is not transcribed prior to metamorphosis; its primary transcript rises rapidly during the first two days of adult life, and remains close to maximal until day 5. Subsequently its level rapidly declines, ultimately reaching values of less than 0.02% of the maximal level. The correlation between primary transcript, mRNA and FABP content were analyzed by modeling transcription, translation and degradation with computer modeling software. The computer simulation is in good agreement with the experimentally obtained data, suggesting that the control of FABP expression in locust flight muscle occurs predominantly at the level of transcription initiation. (C) 1998 Elsevier Science Ltd. All rights reserved.
2.Haunerland, NH. (1997) Transport and utilization in insect flight muscles.Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology 117: 475-482 Transport and utilization in insect flight muscles
fatty acid oxidation; insect flight metabolism; locust; fatty acid transport; lipase; FABP
In migrating lepidopteran and orthopteran insects, lipid is the preferred fuel for sustained flight activity. Diacylglycerol is delivered by lipophorin to the flight muscle and hydrolyzed to free fatty acid and glycerol. After penetrating the plasma membrane by an unknown mechanism, fatty acids are bound by the intracellular fatty acid binding protein (FABP) and transported through the cytosol. After their conversion to acyl-CoA esters, the fatty acids enter the mitochondrial matrix via the carnitine shuttle for subsequent P-oxidation. This article reviews the current knowledge of lipid metabolism in insect flight muscle, with particular emphasis on the structure and function of FABP and its expression during locust development and flight. (C) 1997 Elsevier Science Inc.
1. Zhang, YY; Haunerland, NH; Punja, ZK. (1996) Chitinase profiles in mature carrot (Daucus carota) roots and purification and characterization of a novel isoform.Physiologia Plantarum 96: 130-138 Chitinase profiles in mature carrot (Daucus carota) roots and purification and characterization of a novel isoform
antifungal; carrot; chitinase; Daucus carota; isoforms; pathogenesis-related protein
The profile of chitinases (EC 3.2.1.14) in mature carrot (Daucus carota L. cv. Eagle) roots was studied. Multiple chitinase bands (8-10) were observed in native and sodium dodecylsulfate-denaturing polyacrylamide gels. The molecular masses of these chitinases were estimated to be from 20 000 to 40 000. One major chitinase was purified and found to be an acidic protein with pI at 4.3 and a molecular mass of 39 500. The optimum pH for enzymatic activity was around 5 and the optimum temperature was 25 degrees C. The enzyme was stable at pHs below 8 and temperatures below 60 degrees C. The protein did not have a chitin-binding domain, but showed some similarity to the amino acid composition of tobacco class I chitinase. The N-terminal amino acid sequence did not resemble any of the described classes of chitinases. This chitinase did not possess lysozyme activity and showed antifungal activity when tested against Trichoderma sp.
