Biomedical subjects

I Lee

Publications and source records attributed to I Lee.

At least 109 records · Page 6Linked to original sources

PubMed1998

Effects of Dispersant Treatment on the Acute Aquatic Toxicity of Petroleum Hydrocarbons

The acute effects of both untreated and dispersant-treated Prudhoe Bay crude oil on the early life-stages of three marine species were investigated. Identification of which water-accommodated fraction (undispersed or chemically dispersed) was considered "more toxic" was dependent on species, time, and endpoint (and by inference, test protocol). Generally, the data showed that at roughly equivalent hydrocarbon concentrations untreated oil solutions resulted in higher initial effects (< 1 h) in mysid and topsmelt tests, whereas dispersed oil solutions elicited higher levels of larval abnormality in abalone tests and higher levels of mortality in mysid tests. While differences in test protocols existed among the species tested, topsmelt were the most sensitive species to untreated oil solutions, with mysids being most sensitive to dispersed oil solutions.

Journal Article↗

PubMed1998

Ultrastructural localization and biochemical characterization of vitronectin in developing rat bone.

This study has used light and electron microscope immunohistochemical and biochemical methods to localize and characterize vitronectin in early bone formation of developing rat mandible with rabbit antimurine vitronectin IgG. Developing jaws of foetuses were collected at embryonic day 15 (day 15) to day 18 from pregnant Wistar rats. After aldehyde fixation, specimens with and without osmium post-fixation were dehydrated and embedded in paraffin, Spurr's resin or LR gold resin for morphological and immunohistochemical examinations. At the light microscope level, in day 15 samples, positive vitronectin immunostaining was observed in small elongated areas of intercellular matrix and osteoblasts. Concomitant with initiation of matrix mineralization at day 16, vitronectin staining was similarly observed in small elongated areas containing intercellular matrix and osteoblasts but not clearly detected in fully mineralized bone matrix. The same staining profile was observed at days 17 and 18. At the ultrastructural level, immunogold particles were clearly detected over unmineralized matrix and cisterns of the rough-surfaced endoplasmic reticulum and the Golgi apparatus of osteoblasts as well as over demineralized bone matrix at day 16-18. In order to assess the presence of vitronectin in the mineral phase, mineral-binding bone proteins were extracted from fresh day 18 specimens using a three-step technique: 4M guanidine HCl (G1 extract), aqueous EDTA without guanidine HCl (E extract), followed by guanidine HCl. Subsequent Western blot analysis of sodium dodecyl sulphate (SDS)-polyacrylamide gel electrophoresis revealed that the antibodies produced only a single band at an M(r) of approximately 73000 in both G1 and E extracts, indicating the presence of vitronectin in the mineralized bone matrix. These results indicate that, at the onset of bone formation, osteoblasts synthesize and release vitronectin, which is subsequently incorporated into the bone matrix and becomes a specific component of bone tissues. The observation of vitronectin in these critical stages of bone formation suggests that it may be involved in the regulation of bone formation.

Animals↗

PubMed1998

A novel tumor necrosis factor-alpha mutant with significantly enhanced cytotoxicity and receptor binding affinity.

A novel tumor necrosis factor-alpha mutant (mutant M3), in which Ser and Tyr at positions 52 and 56 were substituted by Ile and Phe, respectively, along with deletion of 7 N-terminal amino acids, was prepared and its biological activities were investigated. The mutant exhibited a 14- to 24-fold increase in the cytotoxicity relative to the wild-type TNF on various cancer cell lines. The binding affinity of the mutant to TNF-R55 and TNF-R75 receptors was over 10-fold higher than that of the wild-type. TNF-alpha and the mutant show similar CD spectra in the far-UV region, indicating that the overall structure was not influenced by the mutations. The production of highly potent TNF-alpha mutant utilizing increase of hydrophobicity in the region 52-56 may provide a structural basis for a design of optimized TNF-alpha as a therapeutic purpose.

Animals↗

PubMed1998

Characteristic immunolocalization of Ku protein as nuclear matrix.

Two hybridoma clones, NMB1 and NML90, were established using nuclear matrix proteins from normal human thymi or malignant lymphoma as immunogens. They reacted with human Ku70 and Ku80, respectively, by immunoblotting. When HeLa cell nuclear proteins were fractionated and applied to immunoblotting, both Ku70 and Ku80 were detected in the nuclear matrix as well as the soluble nuclear protein fractions. By confocal scanning microscopy, the immunoreactivity of Ku70 and Ku80 was localized to distinct nucleoplasmic fibrillar network and fine granules in the interphase cell nuclei. When HeLa cells were fractionated in situ using DNase I and buffers containing 0.25 M (NH4)2SO4 and 2 M NaCl, the nucleoplasmic reticular structure was largely preserved, but granules disappeared. The nucleoplasmic distribution of Ku in the tissue and in cultured cells was distinct from each other. In the adult tissue, it consisted mostly of either distinct curvilinear lines along the nuclear periphery or of tangled, beaded lines throughout the nuclei. When xenotransplants of HeLa cell in Scid mice were examined, the "tissue type" immunolocalization pattern was reproduced consistently. In most fetal tissues, "tissue type" and "cell type" patterns were admixed. Monoclonal antibodies described here are useful tools for studying the structure and function of the nuclear matrix.

Animals↗

PubMed1998

Flowering-time genes modulate the response to LEAFY activity.

Among the genes that control the transition to flowering in Arabidopsis is a large group whose inactivation causes a delay in flowering. It has been difficult to establish different pathways in which the flowering-time genes might act, because mutants with lesions in these genes have very similar phenotypes. Among the putative targets of the flowering-time genes is another group of genes, which control the identity of individual meristems. Overexpression of one of the meristem-identity genes, LEAFY, can cause the precocious generation of flowers and thus early flowering. We have exploited the opposite phenotypes seen in late-flowering mutants and LEAFY overexpressers to clarify the genetic interactions between flowering-time genes and LEAFY. According to epistatic relationships, we can define one class of flowering-time genes that affects primarily the response to LEAFY activity, and another class of genes that affects primarily the transcriptional induction of LEAFY. These observations allow us to expand previously proposed models for the genetic control of flowering time.

Arabidopsis Proteins↗

PubMed1998

Engineered human carboxypeptidase B enzymes that hydrolyse hippuryl-L-glutamic acid: reversed-polarity mutants.

Variants of human pancreatic carboxypeptidase B (HCPB), with specificity for hydrolysis of C-terminal glutamic acid and aspartic acid, were prepared by site-directed mutagenesis of the human gene and expressed in the periplasm of Escherichia coli. By changing residues in the lining of the S1' pocket of the enzyme, it was possible to reverse the substrate specificity to give variants able to hydrolyse prior to C-terminal acidic amino acid residues instead of the normal C-terminal basic residues. This was achieved by mutating Asp253 at the base of the S1' specificity pocket, which normally interacts with the basic side-chain of the substrate, to either Lys or Arg. The resulting enzymes had the desired reversed polarity and enzyme activity was improved significantly with further mutations at residue 251. The [G251T,D253K]HCPB double mutant was 100 times more active against hippuryl-L-glutamic acid (hipp-Glu) as substrate than was the single mutant, [D253K]HCPB. Triple mutants, containing additional changes at Ala248, had improved activity against hipp-Glu substrate when position 251 was Asn. These reversed-polarity mutants of a human enzyme have the potential to be used in antibody-directed enzyme prodrug therapy of cancer.

Alanine↗

PubMed1998

Cartilage rods as a potential material for penile reconstruction.

PURPOSE: Sex assignment is made in patients with ambiguous genitalia, genital trauma or iatrogenic injury after a thorough diagnostic evaluation and careful consultation with the family. In numerous instances a decision is made to rear the child as the female gender due to inadequate genitalia regardless of karyotype. Although a silicone penile prosthesis is accepted treatment in adults who require penile reconstruction, it has not been generally used in the pediatric population, mainly due to associated long-term problems. We determine the feasibility of creating natural penile prostheses of cartilage which, if biocompatible and elastic, may be used in patients who require genital reconstruction. MATERIALS AND METHODS: Cartilage was harvested from the articular surface of calf shoulders. Chondrocytes were isolated, grown and expanded in vitro. Cells were seeded onto preformed cylindrical polyglycolic acid polymer rods 1 cm. in diameter and 3 cm. long at a concentration of 50 x 10(6) chondrocytes per cm.3. A total of 40 polymer scaffolds were implanted in the subcutaneous space of 20 athymic mice. In each mouse 2 implantation sites consisted of a polymer scaffold seeded with chondrocytes and a control (polymer alone). Mice were sacrificed 1, 2, 4 and 6 months after implantation, respectively. Stress relaxation studies to measure biomechanical properties, including compression, tension and bending, were performed on the retrieved structures. Histological analyses were done with hematoxylin and eosin, aldehyde fuchsin-alcian blue and toluidine blue staining. RESULTS: Gross examination revealed well formed, milk-white rod-shaped solid cartilaginous structures the same size as the initial implant. Compression, tension and bending studies demonstrated that the cartilaginous structures were readily elastic and withstood high degrees of pressure. Histochemical analyses showed mature, well formed chondrocytes in all implants. There was no evidence of cartilage formation in the controls. CONCLUSIONS: Chondrocytes seeded on preformed biodegradable polymer structures form cartilage rods. The use of an entirely autologous system composed of biodegradable polymers and chondrocytes precludes an immunological reaction. This technology appears to be useful for the creation of a biocompatible malleable penile prosthesis, which may be useful in children with ambiguous genitalia and patients undergoing penile reconstruction.

Animals↗

PubMed1998

Axial heterogeneity of sodium-bicarbonate cotransporter expression in the rabbit proximal tubule.

It is generally accepted that Na(HCO3)n cotransport is the most important mechanism mediating basolateral bicarbonate efflux in the early proximal tubule. The presence of basolateral Na(HCO3)n cotransport in the late proximal tubule (S3 segment) and in the juxtamedullary S1 and S2 segments has been controversial. The renal sodium-bicarbonate cotransporter (NBC) has been recently cloned from rat (M. F. Romero, M. A. Hediger, E. L. Boulpaep, and W. F. Boron. J. Am. Soc. Nephrol. 7: 1259, 1996), salamander (M. F. Romero, M. A. Hediger, E. L. Boulpaep, and W. F. Boron. Nature 387: 409-413, 1997), and human (C. E. Burnham, H. Amlal, Z. Wang, G. E. Shull, and M. Soleimani. J. Biol. Chem. 272: 19111-19114, 1997). The localization of NBC in the kidney is unknown. The present study was designed to localize NBC mRNA expression in the rabbit proximal tubule. In situ hybridization studies were combined with functional studies of basolateral Na(HCO3)n cotransport in superficial and juxtamedullary S1, S2, and S3 segments of the rabbit proximal tubule. The results demonstrate that NBC mRNA is localized predominantly to the cortex, with less expression in the outer medulla. NBC expression was not detected in the inner medulla. The highest level of NBC mRNA is in the S1 proximal tubule. NBC is expressed at a low levels in the S3 segment, with intermediate expression in the S2 segment. In bicarbonate-buffered solutions, the rate of base efflux mediated by Na(HCO3)n cotransport followed a similar pattern in superficial and juxtamedullary proximal tubule segments, i.e., S1 > S2 > S3. The juxtamedullary S1 segment had the greatest rate of basolateral Na(HCO3)n cotransport and the highest level of NBC expression in the proximal tubule.

Animals↗

PubMed1998

Low expression of the apolipoprotein B mRNA-editing transgene in mice reduces LDL levels but does not cause liver dysplasia or tumors.

Hepatic expression of apolipoprotein (apo) B mRNA-editing enzyme catalytic polypeptide 1 (APOBEC-1) has been proposed as a gene therapy approach for lowering plasma low density lipoprotein (LDL) levels. However, high-level expression of APOBEC-1 in transgenic mouse and rabbit livers causes liver dysplasia and hepatocellular carcinoma. To determine the physiological and pathological effects of low-level hepatic expression of APOBEC-1, we used a 52-kb rat APOBEC-1 genomic clone (RE4) to generate transgenic mice expressing low levels of APOBEC-1 (2 to 5 times those in nontransgenic mice). Liver function, liver histology, editing of apoB mRNA at the normal editing site (C6666), and abnormal editing at multiple sites (hyperediting) in these mice were compared with those in transgenic mice expressing intermediate (I-20) or high (I-28) levels of APOBEC-1 in the liver. Hyperediting of mRNA coding for the novel APOBEC-1 target 1 (NAT1) was also examined. In the high-expressing I-28 line, 50% of the mice had palpable tumors at 15 weeks of age, whereas in the intermediate-expressing I-20 line, 50% of the mice had evidence of liver tumors after 1 year. In contrast, low-expressing RE4 mice had normal liver function and histology and did not develop liver tumors when examined at 3 to 17 months of age. Moreover, hyperediting of apoB and NAT1 mRNA in the liver was robust in the I-20 mice but barely detectable in the RE4 mice. The low-level expression resulted in sufficient APOBEC-1 to edit essentially all apoB mRNA at the normal editing site, virtually eliminating apoB-100 and LDL in the plasma of RE4 mice. When RE4 mice were crossed with human apoB transgenic mice, which possess high plasma LDL concentrations, plasma LDL levels in the offspring were reduced to very low levels. These results indicates that long-term hepatic expression of APOBEC-1 at low levels sufficient to eliminate LDL does not cause apparent liver damage or liver tumors in transgenic mice. RE4 APOBEC-1 transgenic mice should prove valuable for studying the roles of apoB-containing lipoproteins in lipid metabolism and atherosclerosis.

APOBEC-1 Deaminase↗

PubMed1998

The N-terminal portion of parathyroid hormone-related protein mediates the inhibition of apical Na+/H+ exchange in opossum kidney cells.

Parathyroid hormone (PTH) and PTH-related protein (PTHrP) can activate a common receptor in several different cell types. Both PTH and N-terminal PTHrP peptides have been shown to acutely inhibit the apical Na+/H+ exchanger in the renal proximal tubule. In this study, the ability of various PTHrP fragments to inhibit apical Na+/H+ exchange was investigated. In addition, the signal transduction events associated with PTHrP inhibition of apical Na+/H+ exchange in polarized OK-P cells were characterized. Both PTHrP-(1-34)NH2 and recombinant full-length PTHrP-(1-141) inhibited apical Na+/H+ exchange activity by approximately 50%. These changes occurred in close temporal association with significant (threefold) increases in cellular cAMP accumulation. PTHrP-(1-34)NH2 had no effect on intracellular Ca2+, inositol phosphate production, or protein kinase C activity. PTHrP peptides, including PTHrP-(38-64)NH2, PTHrP-(67-86)NH2, PTHrP-(102-107)NH2, and PTHrP-(107-139)NH2, which lack the PTH-like N terminus, had no effect on the antiporter activity or cAMP accumulation. The results demonstrate that the N-terminal portion of the PTHrP molecule is responsible for inhibition of the apical Na+/H+ antiporter in OK-P cells.

Animals↗

PubMed1998

Peripheral blood mononuclear cells express mutated NCCT mRNA in Gitelman's syndrome: evidence for abnormal thiazide-sensitive NaCl cotransport.

Genetic analysis has demonstrated complete linkage between the human thiazide-sensitive sodium chloride cotransporter gene (NCCT or TSC) and Gitelman's syndrome (GS). Several genomic NCCT mutations have been reported. This study was performed to determine whether peripheral blood mononuclear cells (PBMC) express NCCT mRNA and whether defective PBMC NaCl cotransport could be demonstrated in GS. PBMC were isolated from two brothers with GS, their parents, and healthy control subjects. Northern analysis revealed that NCCT mRNA is expressed in PBMC. The sequence of full-length NCCT cDNA amplified from normal PBMC was identical to human renal NCCT cDNA. Two different mutations were detected in the patients' NCCT cDNA (compound heterozygote). In cDNA derived from the patient's maternal allele, exon 24 was deleted, resulting in a premature stop codon (after amino acid 920). cDNA derived from the patient's paternal allele had an additional 119-bp insertion between exons 3 and 4, generating a premature stop codon (after amino acid 187). The patient's genomic DNA had a previously described 5' splice site mutation in intron 24, GGT --> GTT (maternal allele), and a new 3' splice site mutation in intron 3, CAG --> CAA (paternal allele), which resulted in the activation of a nearby cryptic splice site in intron 3. The latter mutation was not present in 300 normal chromosomes. To determine the functional significance of these findings, chlorothiazide-inhibitable 22Na uptake was measured in PBMC from control subjects, the parents, and the patients with GS in the presence of bumetanide. In control PBMC, chlorothiazide inhibited 22Na uptake by approximately 9%. PBMC from the two patients with GS failed to respond to chlorothiazide. These results demonstrate that PBMC can be used for mutational analysis of NCCT mRNA in patients with GS. Furthermore, functional evidence is provided that the underlying cause of GS is defective NCCT NaCl cotransport.

Adult↗

PubMed1998

Immunocytochemical localization and biochemical characterization of large proteoglycans in developing rat bone.

This study used biochemical and light and electron microscopic immunohistochemical methods to localize and characterize large hyaluronate-binding proteoglycans in the developing mandible of fetal rats at embryonic day 15 (Day 15) to Day 18 using a monoclonal antibody (MAb) 5D5. This antibody is derived from bovine sclera and specifically recognizes the core protein of large proteoglycan such as versican, neurocan and brevican, but not that of aggrecan. At the light microscopic level, MAb 5D5 moderately stained the extracellular matrices among osteoblasts at the centers of ossification in Day 15 mandible specimens. Weaker staining was observed in osteoblasts, whereas Meckel's cartilage lacked staining. Ultrastructural immunocytochemistry showed the presence of immunogold particles over unmineralized matrices among osteoblasts and their intracellular organelles. In Day 16 to 18 specimens, bone nodules were recognized in LR gold sections before immunostaining, but, after immunostaining, consistently appeared devoid of mineral crystals and were seen as a demineralized structure that had an electron dense periphery within which fine filamentous and granular material were present. The appearance of these structures was created by the demineralization of thin sections on grids during immunostaining. Specific immunogold staining was clearly seen over the demineralized structures corresponding to bone nodules. The majority of immunogold particles tended to localize inside of the structures. Bone proteins were extracted from fresh, Day 18 specimens with a three-step technique: 4 M guanidine HCl (GdnCl,-extract), aqueous EDTA without GdnCl (E-extract), followed by GdnCl. Western blot analysis of SDS-polyacrylamide gel electrophoresis after chondroitinase ABC digestion, showed that G1-extract gave a 5D5 reactive band greater than 400 kDa, whereas E-extract produced two major reactive populations of small molecular size with core proteins approximately 63 and 74 kDa. These results indicate that the large proteoglycan having smaller molecular weight is preferentially localized to bone nodules and may correlate with bone matrix mineralization.

Animals↗

PubMed1998

A novel nuclear substructure, ND10: distribution in normal and neoplastic human tissues.

ND10 are recently characterized nuclear domains that are composed of 0.5 microm sized, precisely circumscribed dots in cultured human cell lines. To investigate the distribution and number of ND10 on various types of normal and neoplastic human tissues, we carried out immunostaining and immunoprecipitation analyses with monoclonal antibodies 138 and 1150. The number of ND10 varied from 1 to 10 or more in various tissues as did their size. ND10 were diffusely located in early embryonic and normal tissues, except for the exocrine and endocrine cells of the pancreas and for hepatocytes. In normal squamous mucosa, basal cells had more ND10 than did differentiated superficial squamous cells. The number and size of ND10 were markedly increased in malignant neoplasms but were similar in benign tumors and corresponding normal tissues. Sex hormone-related normal tissues, such as the endometrium or myometrium, and neoplasms strongly stained for ND10. The distribution pattern of ND10 in human tissues indicates that they are conserved nuclear substructures that are closely associated with cellular differentiation, hormonal stimulation, and oncogenesis.

Digestive System↗

PubMed1997

The existence of substance P in Purkinje cells in cerebellum of the gerbil.

We performed this study to identify the characteristics of substance P localization in gerbil cerebellum by immunohistochemistry. Substance P immunoreactivity was present in Purkinje cells of cerebellar cortex but not in other structures. It is suggested that the cerebellar mechanism of gerbil may be different from that of other species.

Animals↗

PubMed1997

Evaluation of cellular uptake, tumor retention, radiation response, and tumor pathophysiology in experimental solid tumors after an intratumoral infusion of colloidal 32P.

BACKGROUND: Order et al. successfully deposited > 85% of radionuclides in pancreatic carcinoma patients using an intratumoral (i. t.) infusion of macroaggregated albumin (MAA) prior to the i. t. infusion of colloidal 32P (32P-CP), apparently due to MAA-induced transient blockade. However, no studies regarding physiologic response and the inhibitory tumor growth of solid tumors after treatment with 32P-CP with or without MAA have been performed. METHODS: First, the authors determined the cellular uptake of 32P-CP in 10 cell lines. They then evaluated the relationship between tumor retention and radiation response in vivo. Third, they evaluated the changes in physiologic parameters such as tumor interstitial fluid pressure (TIFP) and tumor blood flow (TBF) after an i. t. infusion of MAA. RESULTS: In the plateau growth phase of both H4IIE and LS174T tumors, maximal uptake occurred at approximately 100 minutes after treatment with 10 microCi of 32P-CP, but uptake in LS174T was approximately 2 times higher than that in H4IIE. In animal experiments, 32P-CP alone significantly inhibited the tumor growth of H4IIE. However, additional i. t. infusion of MAA did not improve the growth delay induced by 32P-CP, mainly due to insignificant differences in retention of radioactivity when using 32P-CP with or without MAA. It was speculated that when the outflow of the tumor vasculature was obstructed by clamping (or an i. t. infusion of MAA), it would reduce TBF and increase TIFP. However, neither increased TIFP nor decreased TBF was observed when MAA was given i. t. to tumors. CONCLUSIONS: The authors concluded that an MAA-induced transient blockade of tumor vasculature after an i. t. infusion of MAA did not occur in experimental solid tumors.

Albumins↗

PubMed1997

Biphasic transitions of a hairpin hexanucleotide triplex DNA.

The conformational transitions (helix-coil transitions) of three hairpin triple helices, models 5'-(A-G)(3) + 5'-(T-C)(3)-T(4)-((br)C-T)(3) [CY], 5'-(A-G)(3) + 5'-(T-(br)C)(3)-T(4)-(C-T)(3) [YC] and 5'-(A-G)(3) + 5'-(T-(br)C)(3)-T(4)-((br)C-T)(3) [YY], are characterized in this work by UV spectroscopy. Melting of these triplexes is biphasic, and the profiles are used to obtain the thermodynamic parameters. The thermodynamic properties of the hairpin triplex are T(m) = 19.45 degrees C and DeltaH(vH) = 293.12 kJ mol(-1) for CY, T(m) = 22.85 degrees C and DeltaH(vH) = 256.63 kJ mol(-1) for YC and T(m) = 28.47 degrees C and DeltaH(vH) = 234.68 kJ mol(-1) for YY at pH 4.4. Those of the duplex are T(m) = 30.50 degrees C and DeltaH(vH) = 427.09 kJ mol(-1) for CY, T(m) = 32.96 degrees C and DeltaH(vH) = 374.47 kJ mol(-1) for YC and T(m) = 33.24 degrees C and DeltaH(vH) = 329.67 kJ mol(-1) for YY at pH 4.4. The distinct transitions of triplex to duplex and duplex to single strands are analyzed using the nearest-neighbor Ising model. Electrostatic effects on each conformation are also analyzed.

Journal Article↗

PubMed1997

Diversity among clinical isolates of Helicobacter pylori in Korea.

Eleven strains of Helicobacter pylori were isolated from biopsy specimens obtained at the A-San Medical Center from December, 1995 to February, 1996. Every H. pylori positive patient was diagnosed to have chronic, erosive, or mild superficial gastritis. To determine the diversity in clinical isolates, the following were studied: total protein profile, plasmid profile, presence of cagA, and variation in DNA sequence. Protein profiles of nine isolates were similar to each other while two isolates had a 35 kDa protein which did not appear in others. The presence of cagA was detected with PCR in seven isolates (63%). Among eleven isolates, seven (63%) carried plasmids. Each isolate showed a big diversity with a PCR-based randomly amplified polymorphic DNA method. Even though all H. pylori isolates used in this study were isolated from gastritis patients at the same hospital, their molecular and biological characteristics were quite different from another showing a big diversity in H. pylori.

Antigens, Bacterial↗

PubMed1997

The subunit structure of cytochrome-c oxidase from tuna heart and liver.

Cytochrome-c oxidase was isolated from tuna liver and heart, and the subunit composition was analysed by SDS/PAGE by two separation systems. Two additional subunits of the enzyme complex were immunoprecipitated from solubilized mitochondria with an antibody against bovine subunit IV. The N-terminal and internal amino acid sequences of all nuclear-coded subunits were determined after blotting onto poly(vinylidene difluoride) membranes or by tryptic hydrolysis of gel bands and HPLC separation of peptides, respectively. 13 subunits were identified with isoforms for subunits Va, VIc, VIIb and VIII. The isoforms for subunits Va and VIIb are found in liver and heart, isoforms for subunit VIc only in heart, and isoforms for subunit VIII only in liver. Isoforms for subunits Va, VIc and VIIb have not been described in other species. The postulated mechanism of thermogenesis in mammals, based on decreased H+/e- stoichiometry at high ATP/ADP ratios due to binding of ATP to the heart-type subunit VIa [Frank, V. & Kadenbach, B. (1996) FEBS Lett. 382, 121-124], appears not to occur in tuna, because no isoforms of subunit VIa were found.

Amino Acid Sequence↗