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Replacement of C305 in Heart/Muscle-Type Isozyme of Human Carnitine Palmitoyltransferase I with Aspartic Acid and Other Amino Acids

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Abstract

Liver- and heart/muscle-type isozymes of human carnitine palmitoyltransferase I (L- and M-CPTI, respectively) show a certain similarity in their amino acid sequences, and mutation studies on the conserved amino acids between these two isozymes often show essentially the same effects on their enzymatic properties. Earlier mutation studies on C305 in human M-CPTI and its counterpart residue, C304, in human L-CPTI showed distinct effects of the mutations, especially in the aspect of enzyme stability; however, simple comparison of these effects on the conserved Cys residue between L- and M-CPTI was difficult, because these studies were carried out using different expression systems and distinct amino acids as replacements. In the present study, we carried out mutation studies on the C305 in human M-CPTI using COS cells for the expression system. Our results showed that C305 was replaceable with aspartic acid but that substitution with other amino acids caused both loss of function and reduced expression.

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References

  • Brown NF, Mullur RS, Subramanian I, Esser V, Bennett MJ, Saudubray JM, Feigenbaum AS, Kobari JA, Macleod PM, McGarry JD, Cohen JC (2001) Molecular characterization of L-CPT I deficiency in six patients: insights into function of the native enzyme. J Lipid Res 42:1134–1142

    CAS  PubMed  Google Scholar 

  • Dai J, Zhu H, Woldegiorgis G (2003) Leucine-764 near the extreme C-terminal end of carnitine palmitoyltransferase I is important for activity. Biochem Biophys Res Commun 301:758–763

    Article  CAS  PubMed  Google Scholar 

  • Demaugre F, Bonnefont JP, Colonna M, Cepanec C, Leroux JP, Saudubray JM (1991) Infantile form of carnitine palmitoyltransferase II deficiency with hepatomuscular symptoms and sudden death. Physiopathological approach to carnitine palmitoyltransferase II deficiencies. J Clin Invest 87:859–864

    Article  CAS  PubMed  Google Scholar 

  • Esser V, Britton CH, Weis BC, Foster DW, McGarry JD (1993) Cloning, sequencing, and expression of a cDNA encoding rat liver carnitine palmitoyltransferase I. Direct evidence that a single polypeptide is involved in inhibitor interaction and catalytic function. J Biol Chem 268:5817–5822

    CAS  PubMed  Google Scholar 

  • Hashimoto M, Shinohara Y, Majima E, Hatanaka T, Yamazaki N, Terada H (1999) Expression of the bovine heart mitochondrial ADP/ATP carrier in yeast mitochondria: significantly enhanced expression by replacement of the N-terminal region of the bovine carrier by the corresponding regions of the yeast carriers. Biochim Biophys Acta 1409:113–124

    Article  CAS  PubMed  Google Scholar 

  • Ho SN, Hunt HD, Horton RM, Pullen JK, Pease LR (1989) Site-directed mutagenesis by overlap extension using the polymerase chain reaction. Gene 77:51–59

    Article  CAS  PubMed  Google Scholar 

  • IJlst L, Mandel H, Oostheim W, Ruiter JP, Gutman A, Wanders RJ (1998) Molecular basis of hepatic carnitine palmitoyltransferase I deficiency. J Clin Invest 102:527–531

    Article  CAS  PubMed  Google Scholar 

  • Liu H, Zheng G, Treber M, Dai J, Woldegiorgis G (2005) Cysteine-scanning mutagenesis of muscle carnitine palmitoyltransferase I reveals a single cysteine residue (Cys-305) is important for catalysis. J Biol Chem 280:4524–4531

    Article  CAS  PubMed  Google Scholar 

  • McGarry JD, Brown NF (1997) The mitochondrial carnitine palmitoyltransferase system. From concept to molecular analysis. Eur J Biochem 244:1–14

    Article  CAS  PubMed  Google Scholar 

  • Niwa H, Yamamura K, Miyazaki J (1991) Efficient selection for high-expression transfectants with a novel eukaryotic vector. Gene 108:193–199

    Article  CAS  PubMed  Google Scholar 

  • Pan Y, Cohen I, Guillerault F, Feve B, Girard J, Prip-Buus C (2002) The extreme C terminus of rat liver carnitine palmitoyltransferase I is not involved in malonyl-CoA sensitivity but in initial protein folding. J Biol Chem 277:47184–47189

    Article  CAS  PubMed  Google Scholar 

  • Price N, van der Leij F, Jackson V, Corstorphine C, Thomson R, Sorensen A, Zammit V (2002) A novel brain-expressed protein related to carnitine palmitoyltransferase I. Genomics 80:433–442

    Article  CAS  PubMed  Google Scholar 

  • Shi J, Zhu H, Arvidson DN, Woldegiorgis G (1999) A single amino acid change (substitution of glutamate 3 with alanine) in the N-terminal region of rat liver carnitine palmitoyltransferase I abolishes malonyl-CoA inhibition and high affinity binding. J Biol Chem 274:9421–9426

    Article  CAS  PubMed  Google Scholar 

  • Yamazaki N, Shinohara Y, Shima A, Terada H (1995) High expression of a novel carnitine palmitoyltransferase I like protein in rat brown adipose tissue and heart: isolation and characterization of its cDNA clone. FEBS Lett 363:41–45

    Article  CAS  PubMed  Google Scholar 

  • Yamazaki N, Shinohara Y, Shima A, Yamanaka Y, Terada H (1996) Isolation and characterization of cDNA and genomic clones encoding human muscle type carnitine palmitoyltransferase I. Biochim Biophys Acta 1307:157–161

    PubMed  Google Scholar 

  • Yamazaki N, Yamanaka Y, Hashimoto Y, Shinohara Y, Shima A, Terada H (1997) Structural features of the gene encoding human muscle type carnitine palmitoyltransferase I. FEBS Lett 409:401–406

    Article  CAS  PubMed  Google Scholar 

  • Yamazaki N, Matsuo T, Kurata M, Suzuki M, Fujiwaki T, Yamaguchi S, Terada H, Shinohara Y (2008) Substitutions of three amino acids in human heart/muscle type carnitine palmitoyltransferase I caused by single nucleotide polymorphisms. Biochem Genet 46:54–63

    Article  CAS  PubMed  Google Scholar 

  • Zhu H, Shi J, Treber M, Dai J, Arvidson DN, Woldegiorgis G (2003) Substitution of glutamate-3, valine-19, leucine-23, and serine-24 with alanine in the N-terminal region of human heart muscle carnitine palmitoyltransferase I abolishes malonyl CoA inhibition and binding. Arch Biochem Biophys 413:67–74

    Article  CAS  PubMed  Google Scholar 

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Correspondence to Yasuo Shinohara.

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Matsuo, T., Yamamoto, A., Yamamoto, T. et al. Replacement of C305 in Heart/Muscle-Type Isozyme of Human Carnitine Palmitoyltransferase I with Aspartic Acid and Other Amino Acids. Biochem Genet 48, 193–201 (2010). https://doi.org/10.1007/s10528-009-9301-z

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  • DOI: https://doi.org/10.1007/s10528-009-9301-z

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