Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1994 Nov 15;304(Pt 1):31–34. doi: 10.1042/bj3040031

A stress-regulated protein, GRP58, a member of thioredoxin superfamily, is a carnitine palmitoyltransferase isoenzyme.

M S Murthy 1, S V Pande 1
PMCID: PMC1137447  PMID: 7998951

Abstract

We recently noted the association of carnitine palmitoyltransferase (CPT) activity with a 54 kDa microsomal protein [Murthy and Pande (1993) Mol. Cell Biochem. 122, 133-138] that, based on amino-acid-sequence identity, seemed to be the protein previously described as a 'glucose-regulated protein-58' (GRP58), phosphoinositide-specific phospholipase C, hormone-induced protein-70, endoplasmic-reticulum protein-61 (ERp61), protein disulphide-isomerase, thiol protease, a protein affected in halothane anaesthesia and one that affects renal-tubular functions and the transcriptional activation of the interferon-alpha inducible genes. To ascertain the catalytic identity of this protein unambiguously, we have expressed the corresponding cDNA transiently and stably in human kidney 293 cells as well as in HeLa cells. In each case we found that expression led to an increase in assayable and immunoreactive 54 kDa CPT activity, whereas the protein disulphide-isomerase activity was not increased. In vitro expression in a cell-free transcription and translation system led to the synthesis of a approximately 57 kDa (precursor) protein that was processed to a approximately 54 kDa (mature) protein when microsomes were present; in both these experiments again a large increase in CPT activity was seen. Thus the present data provide compelling evidence that the 54 kDa protein in question is a CPT isoenzyme. It remains to be seen now how the ability of this protein to interconvert acyl-CoA and acylcarnitine would relate to the diverse functions indicated for this protein in vivo.

Full text

PDF
31

Images in this article

33Figure 1
on p.33
33Figure 2
on p.33

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Altieri F., Maras B., Eufemi M., Ferraro A., Turano C. Purification of a 57kDa nuclear matrix protein associated with thiol:protein-disulfide oxidoreductase and phospholipase C activities. Biochem Biophys Res Commun. 1993 Aug 16;194(3):992–1000. doi: 10.1006/bbrc.1993.1919. [DOI] [PubMed] [Google Scholar]
  2. Arion W. J. Measurement of intactness of rat liver endoplasmic reticulum. Methods Enzymol. 1989;174:58–67. doi: 10.1016/0076-6879(89)74010-6. [DOI] [PubMed] [Google Scholar]
  3. Bennett C. F., Balcarek J. M., Varrichio A., Crooke S. T. Molecular cloning and complete amino-acid sequence of form-I phosphoinositide-specific phospholipase C. Nature. 1988 Jul 21;334(6179):268–270. doi: 10.1038/334268a0. [DOI] [PubMed] [Google Scholar]
  4. Bhuiyan A. K., Pande S. V. One-step synthesis of radioactive acyl-CoA and acylcarnitines using rat liver mitochondrial outer membrane as enzyme source. Lipids. 1992 May;27(5):392–395. doi: 10.1007/BF02536155. [DOI] [PubMed] [Google Scholar]
  5. Brown N. F., Sen A., Soltis D. A., Jones B., Foster D. W., McGarry J. D. Expression of precursor and mature carnitine palmitoyltransferase II in Escherichia coli and in vitro: differential behaviour of rat and human isoforms. Biochem J. 1993 Aug 15;294(Pt 1):79–86. doi: 10.1042/bj2940079. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Camp L. A., Hofmann S. L. Purification and properties of a palmitoyl-protein thioesterase that cleaves palmitate from H-Ras. J Biol Chem. 1993 Oct 25;268(30):22566–22574. [PubMed] [Google Scholar]
  7. DiRusso C. C., Heimert T. L., Metzger A. K. Characterization of FadR, a global transcriptional regulator of fatty acid metabolism in Escherichia coli. Interaction with the fadB promoter is prevented by long chain fatty acyl coenzyme A. J Biol Chem. 1992 Apr 25;267(12):8685–8691. [PubMed] [Google Scholar]
  8. Henry M. F., Cronan J. E., Jr A new mechanism of transcriptional regulation: release of an activator triggered by small molecule binding. Cell. 1992 Aug 21;70(4):671–679. doi: 10.1016/0092-8674(92)90435-f. [DOI] [PubMed] [Google Scholar]
  9. Johnson E., Henzel W., Deisseroth A. An isoform of protein disulfide isomerase isolated from chronic myelogenous leukemia cells alters complex formation between nuclear proteins and regulatory regions of interferon-inducible genes. J Biol Chem. 1992 Jul 15;267(20):14412–14417. [PubMed] [Google Scholar]
  10. Kaplitt M. G., Kleopoulos S. P., Pfaff D. W., Mobbs C. V. Estrogen increases HIP-70/PLC-alpha messenger ribonucleic acid in the rat uterus and hypothalamus. Endocrinology. 1993 Jul;133(1):99–104. doi: 10.1210/endo.133.1.8319593. [DOI] [PubMed] [Google Scholar]
  11. Lee A. S. Mammalian stress response: induction of the glucose-regulated protein family. Curr Opin Cell Biol. 1992 Apr;4(2):267–273. doi: 10.1016/0955-0674(92)90042-b. [DOI] [PubMed] [Google Scholar]
  12. Lee A. S. The accumulation of three specific proteins related to glucose-regulated proteins in a temperature-sensitive hamster mutant cell line K12. J Cell Physiol. 1981 Jan;106(1):119–125. doi: 10.1002/jcp.1041060113. [DOI] [PubMed] [Google Scholar]
  13. Lewis M. J., Mazzarella R. A., Green M. Structure and assembly of the endoplasmic reticulum: biosynthesis and intracellular sorting of ERp61, ERp59, and ERp49, three protein components of murine endoplasmic reticulum. Arch Biochem Biophys. 1986 Mar;245(2):389–403. doi: 10.1016/0003-9861(86)90230-4. [DOI] [PubMed] [Google Scholar]
  14. Mader S., White J. H. A steroid-inducible promoter for the controlled overexpression of cloned genes in eukaryotic cells. Proc Natl Acad Sci U S A. 1993 Jun 15;90(12):5603–5607. doi: 10.1073/pnas.90.12.5603. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Martin J. L., Pumford N. R., LaRosa A. C., Martin B. M., Gonzaga H. M., Beaven M. A., Pohl L. R. A metabolite of halothane covalently binds to an endoplasmic reticulum protein that is highly homologous to phosphatidylinositol-specific phospholipase C-alpha but has no activity. Biochem Biophys Res Commun. 1991 Jul 31;178(2):679–685. doi: 10.1016/0006-291x(91)90161-y. [DOI] [PubMed] [Google Scholar]
  16. Mazzarella R. A., Marcus N., Haugejorden S. M., Balcarek J. M., Baldassare J. J., Roy B., Li L. J., Lee A. S., Green M. Erp61 is GRP58, a stress-inducible luminal endoplasmic reticulum protein, but is devoid of phosphatidylinositide-specific phospholipase C activity. Arch Biochem Biophys. 1994 Feb 1;308(2):454–460. doi: 10.1006/abbi.1994.1064. [DOI] [PubMed] [Google Scholar]
  17. Mazzarella R. A., Srinivasan M., Haugejorden S. M., Green M. ERp72, an abundant luminal endoplasmic reticulum protein, contains three copies of the active site sequences of protein disulfide isomerase. J Biol Chem. 1990 Jan 15;265(2):1094–1101. [PubMed] [Google Scholar]
  18. Mobbs C. V., Fink G., Pfaff D. W. HIP-70: a protein induced by estrogen in the brain and LH-RH in the pituitary. Science. 1990 Mar 23;247(4949 Pt 1):1477–1479. doi: 10.1126/science.247.4949.1477. [DOI] [PubMed] [Google Scholar]
  19. Mobbs C. V., Fink G., Pfaff D. W. HIP-70: an isoform of phosphoinositol-specific phospholipase C-alpha. Science. 1990 Aug 3;249(4968):566–567. doi: 10.1126/science.2382136. [DOI] [PubMed] [Google Scholar]
  20. Murthy M. S., Bieber L. L. Purification of the medium-chain/long-chain (COT/CPT) carnitine acyltransferase of rat liver microsomes. Protein Expr Purif. 1992 Feb;3(1):75–79. doi: 10.1016/1046-5928(92)90059-6. [DOI] [PubMed] [Google Scholar]
  21. Murthy M. S., Pande S. V. Carnitine medium/long chain acyltransferase of microsomes seems to be the previously cloned approximately 54 kDa protein of unknown function. Mol Cell Biochem. 1993 May 26;122(2):133–138. doi: 10.1007/BF01076097. [DOI] [PubMed] [Google Scholar]
  22. Nigam S. K., Goldberg A. L., Ho S., Rohde M. F., Bush K. T., Sherman MYu A set of endoplasmic reticulum proteins possessing properties of molecular chaperones includes Ca(2+)-binding proteins and members of the thioredoxin superfamily. J Biol Chem. 1994 Jan 21;269(3):1744–1749. [PubMed] [Google Scholar]
  23. Ohtani H., Wakui H., Ishino T., Komatsuda A., Miura A. B. An isoform of protein disulfide isomerase is expressed in the developing acrosome of spermatids during rat spermiogenesis and is transported into the nucleus of mature spermatids and epididymal spermatozoa. Histochemistry. 1993 Dec;100(6):423–429. doi: 10.1007/BF00267822. [DOI] [PubMed] [Google Scholar]
  24. Roncero C., Goodridge A. G. Hexanoate and octanoate inhibit transcription of the malic enzyme and fatty acid synthase genes in chick embryo hepatocytes in culture. J Biol Chem. 1992 Jul 25;267(21):14918–14927. [PubMed] [Google Scholar]
  25. Shepherd J. C., Schumacher T. N., Ashton-Rickardt P. G., Imaeda S., Ploegh H. L., Janeway C. A., Jr, Tonegawa S. TAP1-dependent peptide translocation in vitro is ATP dependent and peptide selective. Cell. 1993 Aug 13;74(3):577–584. doi: 10.1016/0092-8674(93)80058-m. [DOI] [PubMed] [Google Scholar]
  26. Srivastava S. P., Chen N. Q., Liu Y. X., Holtzman J. L. Purification and characterization of a new isozyme of thiol:protein-disulfide oxidoreductase from rat hepatic microsomes. Relationship of this isozyme to cytosolic phosphatidylinositol-specific phospholipase C form 1A. J Biol Chem. 1991 Oct 25;266(30):20337–20344. [PubMed] [Google Scholar]
  27. Srivastava S. P., Fuchs J. A., Holtzman J. L. The reported cDNA sequence for phospholipase C alpha encodes protein disulfide isomerase, isozyme Q-2 and not phospholipase-C. Biochem Biophys Res Commun. 1993 Jun 30;193(3):971–978. doi: 10.1006/bbrc.1993.1720. [DOI] [PubMed] [Google Scholar]
  28. Wakui H., Komatsuda A., Ishino T., Imai H., Kobayashi R., Nakamoto Y., Miura A. B. Localization of phosphoinositide-specific phospholipase C-alpha in porcine kidney. Kidney Int. 1992 Oct;42(4):888–895. doi: 10.1038/ki.1992.365. [DOI] [PubMed] [Google Scholar]
  29. Walter P., Blobel G. Preparation of microsomal membranes for cotranslational protein translocation. Methods Enzymol. 1983;96:84–93. doi: 10.1016/s0076-6879(83)96010-x. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES