Abstract
We report the cloning and expression of a cDNA encoding human sterol carrier protein 2 (SCP2). The 1.3-kilobase (kb) cDNA contains an open reading frame which encompasses a 143-amino acid sequence which is 89% identical to the rat SCP2 amino acid sequence. The deduced amino acid sequence of the polypeptide reveals a 20-residue amino-terminal leader sequence in front of the mature polypeptide, which contains a carboxyl-terminal tripeptide (Ala-Lys-Leu) related to the peroxisome targeting sequence. The expressed cDNA in COS-7 cells yields a 15.3-kDa polypeptide and increased amounts of a 13.2-kDa polypeptide, both reacting with a specific rabbit antiserum to rat liver SCP2. The cDNA insert hybridizes with 3.2- and 1.8-kb mRNA species in human liver poly(A)+ RNA. In human fibroblasts and placenta the 1.8-kb mRNA was most abundant. Southern blot analysis suggests either that there are multiple copies of the SCP2 gene in the human genome or that the SCP2 gene is very large. Coexpression of the SCP2 cDNA with expression vectors for cholesterol side-chain cleavage enzyme and adrenodoxin resulted in a 2.5-fold enhancement of progestin synthesis over that obtained with expression of the steroidogenic enzyme system alone. These findings are concordant with the notion that SCP2 plays a role in regulating steroidogenesis, among other possible functions.
Full text
PDFImages in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Andersson S., Davis D. L., Dahlbäck H., Jörnvall H., Russell D. W. Cloning, structure, and expression of the mitochondrial cytochrome P-450 sterol 26-hydroxylase, a bile acid biosynthetic enzyme. J Biol Chem. 1989 May 15;264(14):8222–8229. [PubMed] [Google Scholar]
- Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
- 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]
- Chanderbhan R., Tanaka T., Strauss J. F., Irwin D., Noland B. J., Scallen T. J., Vahouny G. V. Evidence for sterol carrier protein2-like activity in hepatic, adrenal and ovarian cytosol. Biochem Biophys Res Commun. 1983 Dec 28;117(3):702–709. doi: 10.1016/0006-291x(83)91654-6. [DOI] [PubMed] [Google Scholar]
- Crain R. C., Zilversmit D. B. Two nonspecific phospholipid exchange proteins from beef liver. I. Purification and characterization. Biochemistry. 1980 Apr 1;19(7):1433–1439. doi: 10.1021/bi00548a026. [DOI] [PubMed] [Google Scholar]
- 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]
- Gould S. G., Keller G. A., Subramani S. Identification of a peroxisomal targeting signal at the carboxy terminus of firefly luciferase. J Cell Biol. 1987 Dec;105(6 Pt 2):2923–2931. doi: 10.1083/jcb.105.6.2923. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Henikoff S. Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene. 1984 Jun;28(3):351–359. doi: 10.1016/0378-1119(84)90153-7. [DOI] [PubMed] [Google Scholar]
- McNamara B. C., Jefcoate C. R. The role of sterol carrier protein 2 in stimulation of steroidogenesis in rat adrenal mitochondria by adrenal cytosol. Arch Biochem Biophys. 1989 Nov 15;275(1):53–62. doi: 10.1016/0003-9861(89)90349-4. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- 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]
- Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Scallen T. J., Seetharam B., Srikantaiah M. V., Hansbury E., Lewis M. K. Sterol carrier protein hypothesis: requirement for three substrate-specific soluble proteins in liver cholesterol biosynthesis. Life Sci. 1975 Mar 15;16(6):853–873. doi: 10.1016/0024-3205(75)90002-8. [DOI] [PubMed] [Google Scholar]
- Takagi K., Alvarez J. G., Favata M. F., Trzaskos J. M., Strauss J. F., 3rd Control of low density lipoprotein receptor gene promoter activity. Ketoconazole inhibits serum lipoprotein but not oxysterol suppression of gene transcription. J Biol Chem. 1989 Jul 25;264(21):12352–12357. [PubMed] [Google Scholar]
- Toaff M. E., Schleyer H., Strauss J. F., 3rd Metabolism of 25-hydroxycholesterol by rat luteal mitochondria and dispersed cells. Endocrinology. 1982 Dec;111(6):1785–1790. doi: 10.1210/endo-111-6-1785. [DOI] [PubMed] [Google Scholar]
- Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Vahouny G. V., Chanderbhan R., Kharroubi A., Noland B. J., Pastuszyn A., Scallen T. J. Sterol carrier and lipid transfer proteins. Adv Lipid Res. 1987;22:83–113. doi: 10.1016/b978-0-12-024922-0.50007-2. [DOI] [PubMed] [Google Scholar]
- 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]
- Yamamoto R., Kao L. C., McKnight C. E., Strauss J. F., 3rd Cloning and sequence of cDNA for human placental cytokeratin 8. Regulation of the mRNA in trophoblastic cells by cAMP. Mol Endocrinol. 1990 Mar;4(3):370–374. doi: 10.1210/mend-4-3-370. [DOI] [PubMed] [Google Scholar]
- Zuber M. X., Mason J. I., Simpson E. R., Waterman M. R. Simultaneous transfection of COS-1 cells with mitochondrial and microsomal steroid hydroxylases: incorporation of a steroidogenic pathway into nonsteroidogenic cells. Proc Natl Acad Sci U S A. 1988 Feb;85(3):699–703. doi: 10.1073/pnas.85.3.699. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]