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. 1991 May 15;88(10):4338–4342. doi: 10.1073/pnas.88.10.4338

Molecular cloning and deduced amino acid sequence of nonspecific lipid transfer protein (sterol carrier protein 2) of rat liver: a higher molecular mass (60 kDa) protein contains the primary sequence of nonspecific lipid transfer protein as its C-terminal part.

T Mori 1, T Tsukamoto 1, H Mori 1, Y Tashiro 1, Y Fujiki 1
PMCID: PMC51654  PMID: 2034675

Abstract

Two types of cDNA for nonspecific lipid transfer protein (nsLTP), identical to sterol carrier protein 2, of rat liver were cloned; one was 787 base pairs (bp) long containing a 429-bp open reading frame of 143 amino acids, with a mass of 15,303 Da (15-kDa protein). The cDNA from the other type was 1966 bp long, including a 1641-bp open reading frame of 547 amino acids, giving a mass of 59,002 Da (60-kDa protein). The deduced primary sequence for the 15-kDa protein was exactly the same as the published sequence of purified nsLTP, except for an extra N-terminal sequence of 20 amino acids, consistent with the finding that nsLTP is synthesized as a larger precursor and processed to a mature form. The sequence for the 60-kDa protein contained, at the 3' end, the full sequence of the 15-kDa protein, a larger precursor to nsLTP. The 15- and 60-kDa proteins, synthesized in vitro from the respective cDNAs, were both immunoprecipitated by rabbit anti-rat liver nsLTP antibody and comigrated in SDS/PAGE with the proteins made in vitro from total liver RNA. These results shed new light on the dispute among several groups of investigators about the crossreactivity of anti-nsLTP antibody with a higher molecular mass, 60-kDa protein. In Northern blot analysis, two major RNA bands, 0.85 and 2.2 kilobases (kb) long, were detected together with two minor bands of 1.6 and 2.9 kb. The 0.85- and 2.2-kb RNAs most likely encode the 15-and 60-kDa proteins, respectively.

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Selected References

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  1. Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Billheimer J. T., Strehl L. L., Davis G. L., Strauss J. F., 3rd, Davis L. G. Characterization of a cDNA encoding rat sterol carrier protein2. DNA Cell Biol. 1990 Apr;9(3):159–165. doi: 10.1089/dna.1990.9.159. [DOI] [PubMed] [Google Scholar]
  3. Bouillon P., Drischel C., Vergnolle C., Duranton H., Kader J. C. The primary structure of spinach-leaf phospholipid-transfer protein. Eur J Biochem. 1987 Jul 15;166(2):387–391. doi: 10.1111/j.1432-1033.1987.tb13527.x. [DOI] [PubMed] [Google Scholar]
  4. Bout A., Teunissen Y., Hashimoto T., Benne R., Tager J. M. Nucleotide sequence of human peroxisomal 3-oxoacyl-CoA thiolase. Nucleic Acids Res. 1988 Nov 11;16(21):10369–10369. doi: 10.1093/nar/16.21.10369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chanderbhan R. F., Kharroubi A. T., Noland B. J., Scallen T. J., Vahouny G. V. Sterol carrier protein2: further evidence for its role in adrenal steroidogenesis. Endocr Res. 1986;12(4):351–370. doi: 10.3109/07435808609035445. [DOI] [PubMed] [Google Scholar]
  6. Chen C., Okayama H. High-efficiency transformation of mammalian cells by plasmid DNA. Mol Cell Biol. 1987 Aug;7(8):2745–2752. doi: 10.1128/mcb.7.8.2745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fujiki Y., Tsuneoka M., Tashiro Y. Biosynthesis of nonspecific lipid transfer protein (sterol carrier protein 2) on free polyribosomes as a larger precursor in rat liver. J Biochem. 1989 Dec;106(6):1126–1131. doi: 10.1093/oxfordjournals.jbchem.a122977. [DOI] [PubMed] [Google Scholar]
  8. Gould S. J., Keller G. A., Hosken N., Wilkinson J., Subramani S. A conserved tripeptide sorts proteins to peroxisomes. J Cell Biol. 1989 May;108(5):1657–1664. doi: 10.1083/jcb.108.5.1657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hijikata M., Wen J. K., Osumi T., Hashimoto T. Rat peroxisomal 3-ketoacyl-CoA thiolase gene. Occurrence of two closely related but differentially regulated genes. J Biol Chem. 1990 Mar 15;265(8):4600–4606. [PubMed] [Google Scholar]
  10. Innis M. A., Myambo K. B., Gelfand D. H., Brow M. A. DNA sequencing with Thermus aquaticus DNA polymerase and direct sequencing of polymerase chain reaction-amplified DNA. Proc Natl Acad Sci U S A. 1988 Dec;85(24):9436–9440. doi: 10.1073/pnas.85.24.9436. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Keller G. A., Pazirandeh M., Krisans S. 3-Hydroxy-3-methylglutaryl coenzyme A reductase localization in rat liver peroxisomes and microsomes of control and cholestyramine-treated animals: quantitative biochemical and immunoelectron microscopical analyses. J Cell Biol. 1986 Sep;103(3):875–886. doi: 10.1083/jcb.103.3.875. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Keller G. A., Scallen T. J., Clarke D., Maher P. A., Krisans S. K., Singer S. J. Subcellular localization of sterol carrier protein-2 in rat hepatocytes: its primary localization to peroxisomes. J Cell Biol. 1989 Apr;108(4):1353–1361. doi: 10.1083/jcb.108.4.1353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Krisans S. K., Thompson S. L., Pena L. A., Kok E., Javitt N. B. Bile acid synthesis in rat liver peroxisomes: metabolism of 26-hydroxycholesterol to 3 beta-hydroxy-5-cholenoic acid. J Lipid Res. 1985 Nov;26(11):1324–1332. [PubMed] [Google Scholar]
  14. Miyazawa S., Osumi T., Hashimoto T., Ohno K., Miura S., Fujiki Y. Peroxisome targeting signal of rat liver acyl-coenzyme A oxidase resides at the carboxy terminus. Mol Cell Biol. 1989 Jan;9(1):83–91. doi: 10.1128/mcb.9.1.83. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Morris H. R., Larsen B. S., Billheimer J. T. A mass spectrometric study of the structure of sterol carrier protein SCP2 from rat liver. Biochem Biophys Res Commun. 1988 Jul 15;154(1):476–482. doi: 10.1016/0006-291x(88)90711-5. [DOI] [PubMed] [Google Scholar]
  16. Noland B. J., Arebalo R. E., Hansbury E., Scallen T. J. Purification and properties of sterol carrier protein2. J Biol Chem. 1980 May 10;255(9):4282–4289. [PubMed] [Google Scholar]
  17. Okayama H., Berg P. High-efficiency cloning of full-length cDNA. Mol Cell Biol. 1982 Feb;2(2):161–170. doi: 10.1128/mcb.2.2.161. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ossendorp B. C., van Heusden G. P., Wirtz K. W. The amino acid sequence of rat liver non-specific lipid transfer protein (sterol carrier protein 2) is present in a high molecular weight protein: evidence from cDNA analysis. Biochem Biophys Res Commun. 1990 Apr 30;168(2):631–636. doi: 10.1016/0006-291x(90)92367-9. [DOI] [PubMed] [Google Scholar]
  19. Osumi T., Fujiki Y. Topogenesis of peroxisomal proteins. Bioessays. 1990 May;12(5):217–222. doi: 10.1002/bies.950120505. [DOI] [PubMed] [Google Scholar]
  20. Pastuszyn A., Noland B. J., Bazan J. F., Fletterick R. J., Scallen T. J. Primary sequence and structural analysis of sterol carrier protein 2 from rat liver: homology with immunoglobulins. J Biol Chem. 1987 Sep 25;262(27):13219–13227. [PubMed] [Google Scholar]
  21. Rachubinski R. A., Fujiki Y., Mortensen R. M., Lazarow P. B. Acyl-Coa oxidase and hydratase-dehydrogenase, two enzymes of the peroxisomal beta-oxidation system, are synthesized on free polysomes of clofibrate-treated rat liver. J Cell Biol. 1984 Dec;99(6):2241–2246. doi: 10.1083/jcb.99.6.2241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  23. Seltman H., Diven W., Rizk M., Noland B. J., Chanderbhan R., Scallen T. J., Vahouny G., Sanghvi A. Regulation of bile-acid synthesis. Role of sterol carrier protein 2 in the biosynthesis of 7 alpha-hydroxycholesterol. Biochem J. 1985 Aug 15;230(1):19–24. doi: 10.1042/bj2300019. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Stark M. J. Multicopy expression vectors carrying the lac repressor gene for regulated high-level expression of genes in Escherichia coli. Gene. 1987;51(2-3):255–267. doi: 10.1016/0378-1119(87)90314-3. [DOI] [PubMed] [Google Scholar]
  25. Suzuki Y., Yamaguchi S., Orii T., Tsuneoka M., Tashiro Y. Nonspecific lipid transfer protein (sterol carrier protein-2) defective in patients with deficient peroxisomes. Cell Struct Funct. 1990 Oct;15(5):301–308. doi: 10.1247/csf.15.301. [DOI] [PubMed] [Google Scholar]
  26. Tan H., Okazaki K., Kubota I., Kamiryo T., Utiyama H. A novel peroxisomal nonspecific lipid-transfer protein from Candida tropicalis. Gene structure, purification and possible role in beta-oxidation. Eur J Biochem. 1990 May 31;190(1):107–112. doi: 10.1111/j.1432-1033.1990.tb15552.x. [DOI] [PubMed] [Google Scholar]
  27. Tchang F., This P., Stiefel V., Arondel V., Morch M. D., Pages M., Puigdomenech P., Grellet F., Delseny M., Bouillon P. Phospholipid transfer protein: full-length cDNA and amino acid sequence in maize. Amino acid sequence homologies between plant phospholipid transfer proteins. J Biol Chem. 1988 Nov 15;263(32):16849–16855. [PubMed] [Google Scholar]
  28. Thompson S. L., Burrows R., Laub R. J., Krisans S. K. Cholesterol synthesis in rat liver peroxisomes. Conversion of mevalonic acid to cholesterol. J Biol Chem. 1987 Dec 25;262(36):17420–17425. [PubMed] [Google Scholar]
  29. Trzeciak W. H., Simpson E. R., Scallen T. J., Vahouny G. V., Waterman M. R. Studies on the synthesis of sterol carrier protein-2 in rat adrenocortical cells in monolayer culture. Regulation by ACTH and dibutyryl cyclic 3',5'-AMP. J Biol Chem. 1987 Mar 15;262(8):3713–3717. [PubMed] [Google Scholar]
  30. Tsuneoka M., Yamamoto A., Fujiki Y., Tashiro Y. Nonspecific lipid transfer protein (sterol carrier protein-2) is located in rat liver peroxisomes. J Biochem. 1988 Oct;104(4):560–564. doi: 10.1093/oxfordjournals.jbchem.a122510. [DOI] [PubMed] [Google Scholar]
  31. Van der Krift T. P., Leunissen J., Teerlink T., Van Heusden G. P., Verkleij A. J., Wirtz K. W. Ultrastructural localization of a peroxisomal protein in rat liver using the specific antibody against the non-specific lipid transfer protein (sterol carrier protein 2). Biochim Biophys Acta. 1985 Jan 25;812(2):387–392. doi: 10.1016/0005-2736(85)90313-x. [DOI] [PubMed] [Google Scholar]
  32. Westerman J., Wirtz K. W. The primary structure of the nonspecific lipid transfer protein (sterol carrier protein 2) from bovine liver. Biochem Biophys Res Commun. 1985 Feb 28;127(1):333–338. doi: 10.1016/s0006-291x(85)80163-7. [DOI] [PubMed] [Google Scholar]
  33. van Amerongen A., Helms J. B., van der Krift T. P., Schutgens R. B., Wirtz K. W. Purification of nonspecific lipid transfer protein (sterol carrier protein 2) from human liver and its deficiency in livers from patients with cerebro-hepato-renal (Zellweger) syndrome. Biochim Biophys Acta. 1987 Jun 2;919(2):149–155. doi: 10.1016/0005-2760(87)90201-3. [DOI] [PubMed] [Google Scholar]
  34. van Amerongen A., van Noort M., van Beckhoven J. R., Rommerts F. F., Orly J., Wirtz K. W. The subcellular distribution of the nonspecific lipid transfer protein (sterol carrier protein 2) in rat liver and adrenal gland. Biochim Biophys Acta. 1989 Feb 20;1001(3):243–248. doi: 10.1016/0005-2760(89)90106-9. [DOI] [PubMed] [Google Scholar]
  35. van Heusden G. P., Bos K., Raetz C. R., Wirtz K. W. Chinese hamster ovary cells deficient in peroxisomes lack the nonspecific lipid transfer protein (sterol carrier protein 2). J Biol Chem. 1990 Mar 5;265(7):4105–4110. [PubMed] [Google Scholar]

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