Vacuolar system-associated protein-60: a protein characterized from bovine granulosa and luteal cells that is associated with intracellular vesicles and related to human 80K-H and murine beta-glucosidase II.
Brûlé, Sophie; Rabahi, Flora; Faure, Robertet al.
2000 • In Biology of Reproduction, 62 (3), p. 642 - 654
[en] It has been suggested that proteins of molecular size 56-58 kDa play an important role in bovine ovarian follicular development and oocyte maturation. A polyclonal antibody was raised against a 56- to 58-kDa protein band purified from bovine granulosa cells and was used to screen granulosa or luteal cell cDNA expression libraries. This work resulted in the identification of a cDNA encoding for a protein of 60.1 kDa with a signal peptide of 13 residues. The bovine 60.1-kDa protein shared an overall 86.7% and 81.8% identity with, respectively, the human 80K-H protein and the mouse putative beta subunit of glucosidase II (beta-GII), and was named vacuolar system-associated protein-60 (VASAP-60). Marked differences in sequence identity were noted in a putative molecular adapter domain containing a tandem D and E amino acid stretch flanked by proline-rich sequences presenting the minimal PXXP SH3 motif. VASAP-60 was shown to be unglycosylated using endoglycosidase H treatment and was found mainly in a cellular membrane fraction of bovine corpus luteum. VASAP-60 was localized in a rat hepatic Golgi/endosome fraction and in wheat germ agglutinin (WGA) affinity chromatographic eluates, thereby suggesting the presence of interactions with membrane glycoproteins. A polyclonal antibody was raised against the putative adapter domain of the recombinant VASAP-60; this was shown to recognize a major 88-kDa and two minor 58-kDa and 50-kDa proteins, suggesting that the major 88-kDa protein band represents the complete VASAP-60 protein whereas the 58-kDa and the 50-kDa bands represent its proteolytic fragments. Northern blot analysis demonstrated the presence of a single 2.3-kilobase transcript in all the bovine tissues analyzed with variation in the steady state level between tissues. Immunohistochemical observations showed that VASAP-60 was widely distributed in bovine tissues and was localized in pericytoplasmic and perinuclear membranes. In epithelial cells, the staining presented a basolateral or apical polarity associated with intracellular vacuoles. In conclusion, we have characterized a novel acidic membrane protein, associated with organelles of the vacuolar system, that is widely and histospecifically expressed in bovine tissues. VASAP-60 represents either the bovine ortholog or a new family member of the previously characterized human 80K-H and murine beta-GII proteins. Our results suggest that VASAP-60 presents characteristics of a molecular adaptor protein with functions in membrane-trafficking events.
Disciplines :
Veterinary medicine & animal health
Author, co-author :
Brûlé, Sophie; Centre de recherche en reproduction animale, Faculté de médecine vétérinaire, Université de Montréal, St-Hyacinthe, Québec, Canada J2S 7C6
Rabahi, Flora; Ctr. de Rech. en Repro. Animale, Fac. de Med. Veterinaire, Université de Montréal, St-Hyacinthe, Que. J2S 7C6, Canada
Faure, Robert; U. de Rech. en Pediat., Centre Hospitalier Universitaire, Université Laval, Ste-Foy, Que. G1V 4G2, Canada
Beckers, Jean-François ; Université de Liège - ULiège > Département des sciences fonctionnelles (DSF) > Physiologie de la reproduction
Silversides, David W.; Ctr. de Rech. en Repro. Animale, Fac. de Med. Veterinaire, Université de Montréal, St-Hyacinthe, Que. J2S 7C6, Canada
Lussier, Jacques G.; Ctr. de Rech. en Repro. Animale, Fac. de Med. Veterinaire, Université de Montréal, St-Hyacinthe, Que. J2S 7C6, Canada ; Ctr. de Rech. en Repro. Animale, Fac. de Med. Veterinaire, Université de Montréal, St-Hyacinthe, Que. J2S 7C6, Canada
Language :
English
Title :
Vacuolar system-associated protein-60: a protein characterized from bovine granulosa and luteal cells that is associated with intracellular vesicles and related to human 80K-H and murine beta-glucosidase II.
Publication date :
March 2000
Journal title :
Biology of Reproduction
ISSN :
0006-3363
eISSN :
1529-7268
Publisher :
Society for the Study of Reproduction, United States
Salustri A, Hascall VC, Camaioni A, Yanagishita M. Oocyte-granulosa cell interactions. In: Adashi EY, Leung PCK (eds.), The Ovary. New York: Raven Press; 1993; 209-225.
Wassarman PM, Albertini DF. The mammalian ovum. In: Knobil E, Neill JD (eds.), The Physiology of Reproduction. New York: Raven Press; 1994: 79-122.
Eppig JJ. Mammalian oocyte growth and development in vitro. Mol Reprod Dev 1996; 44:260-273.
Rabahi F, Monniaux D, Pisselet C, Chupin D, Durand P. Qualitative and quantitative changes in protein synthesis of bovine follicular cells during the preovulatory period. Mol Reprod Dev 1991; 30:265-274.
Sakai K, Hirai M, Minoshima S, Kudoh J, Fukuyama R, Shimizu N. Isolation of cDNAs encoding a substrate for protein kinase C: nucleotide sequence and chromosomal mapping of the gene for a human 80K protein. Genomics 1989; 5:309-315.
Hirai M, Shimizu N. Purification of two distinct proteins of approximate M, 80 000 from human epithelial cells and identification as proper substrates for protein kinase C. Biochem J 1990; 270:583-589.
Arendt CW, Ostergaard HL. Identification of the CD45-associated 116-kDa and 80-kDa proteins as the α- and β-subunits of α-glucosidase II. J Biol Chem 1997; 272:13117-13125.
Li YM, Mitsuhashi T, Wojciechowicz D, Shimizu N, Li J, Stitt A, He CJ, Banerjee D, Vlassara H. Molecular identity and cellular distribution of advanced glycation endproduct receptors: relationship of p60 to OST-48 and p90 to 80K-H membrane proteins. Proc Natl Acad Sci USA 1996; 93:11047-11052.
Goh KC, Lim YP, Ong SH, Siak CB, Cao X, Tan YH, Guy GR. Identification of p90, a prominent tyrosine-phosphorylated protein in fibroblast growth factor-stimulated cells, as 80K-H. J Biol Chem 1996; 271:5832-5838.
Kanai M, Göke M, Tsunekawa S, Podolsky DK. Signal transduction pathway of human libroblast growth factor receptor 3. J Biol Chem 1997; 272:6621-6628.
Trombetta ES, Simons JF, Helenius A. Endoplasmic reticulum glucosidase II is composed of a catalytic subunit, conserved from yeast to mammals, and a tightly hound noncatalytic HDEL-containing subunit. J Biol Chem 1996; 44:27509-27516.
Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227:680-685.
Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 1979; 76:4350-4354.
Knudsen KA. Proteins transferred to nitrocellulose for use as immunogens. Anal Biochem 1985; 147:285-288.
Harlow E, Lane D. Antibodies. A Laboratory Manual. New York: Cold Spring Harbor Laboratory Press; 1988.
Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 1997; 162:156-159.
Houde A, Lambert A, Saumande J, Silversides DW, Lussier JG. Structure of the bovine follicle-stimulating hormone receptor complementary DNA and expression in bovine tissues. Mol Reprod Dev 1994; 39:127-135.
Sambrook J, Fritsch EF, Maniatis T. Molecular Cloning. A Laboratory Manual. New York: Cold Spring Harbor Laboratory Press: 1989.
Bairoch A, Bucher P, Hofmann K. The PROSITE database, its status in 1995. Nucleic Acids Res 1996; 24:189-196.
Nakai K, Kanehisa M. A knowledge base for predicting protein localization sites in eukaryotic cells. Genomics 1992; 14:897-911.
Herrin DL, Schmidt GW. Rapid, reversible staining of Northern blots prior to hybridization. Biotechniques 1988; 6:196-200.
Singh L, Jones KW. The use of heparin as a simple cost-effective means of controlling background in nucleic acid hybridization procedures. Nucleic Acids Res 1984; 12:5627-5638.
Simmons KR, Caffrey JL, Phillips JL, Abel JH Jr, Niswender GD. A simple method for preparing suspensions of luteal cells. Proc Soc Exp Biol Med 1976; 152:366-371.
Greenberg ME, Bender TP. Isolation of nuclei by sucrose gradient centrifugation. In: Ausubet FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (eds.), Current Protocols in Molecular Biology. New York: John Wiley & Sons, Inc.; 1994: section 4.10.6.
Freeze HH. Endoglycosidase and glycoamidase release of N-linked oligosaccharides. In: Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (eds.), Current Protocols in Molecular Biology. New York: John Wiley & Sons, Inc.; 1994: section 17.13.1.
Faure R, Baquiran G, Bergcron JJ, Posner BI. The dephosphorylation of insulin and epidermal growth factor receptors. Role of endosome-associated phosphotyrosine phosphatase(s). J Biol Chem 1992; 267: 11215-11221.
Bergeron JJM, Cruz J, Khan MN, Posner BI. Uptake of insulin and other ligands into receptor-rich endocytic components of target cells: the endosomal apparatus. Annu Rev Physiol 1985; 47:383-403.
Khan MN, Baquiran G, Brule C, Burgess J, Foster B, Bergeron JJ, Posner BI. Internalization and activation of the rat liver insulin receptor kinase in vivo. J Biol Chem 1989; 264:12931-12940.
Rabahi F, Brûlé S, Sirois J, Beckers J-F, Silversides DW, Lussier JG. High expression of bovine α glutathione S-transferase (GSTA1, GSTA2) subunits is mainly associated with steroidogenically active cells and regulated by gonadotropins in bovine ovarian follicles. Endocrinology 1999; 140:3507-3517.
Kyte J, Doolittle RF. A simple method for displaying the hydropathic character of a protein. J Mol Biol 1982; 157:105-132.
Kozak M. An analysis of 5′-noncoding sequences from 699 vertebrate messenger RNAs. Nucleic Acids Res 1987; 15:8125-8148.
Heijne GV. Transcending the impenetrable: how proteins come to terms with membranes. Biochim Biophys Acta 1988; 947:307-333.
Flura T, Brada D, Ziak M, Roth J. Expression of a cDNA encoding the glucose trimming enzyme glucosidase II in CHO cells and molecular characterization of the enzyme deficiency in a mutant mouse lymphoma cell line. Glycobiology 1997; 7:617-624.
Hentges A, Bause E. Affinity purification and characterization of glucosidase II from pig liver. J Biol Chem 1997; 378:1031-1038.
Brada D, Dubach UC. Isolation of a homogeneous glucosidase II from pig kidney microsomes. Eur J Biochem 1984; 141:149-156.
Arendt CW, Dawicki W, Ostergaard HL. Alternative splicing of transcripts encoding the α- and β-subunits of mouse glucosidase II in T lymphocytes. Glycobiology 1999; 9:277-283.
Strynadka NCJ, James MNG. Crystal structures of the helix-loop-helix calcium-binding proteins. Annu Rev Biochem 1989; 58:951-998.
Nakayama S, Moncrief ND, Kretsinger RH. Evolution of EF-hand calcium-modulated proteins. II. Domains of several subfamilies have diverse evolutionary histories. J Mol Evol 1992; 34:416-448.
Heizmann CW, Hunziker W. Intracellular calcium-binding proteins: more sites than insights. Trends Biochem Sci 1991; 16:98-103.
Ozawa M, Muramatsu T. Reticulocalbin, a novel endoplasmic reticulum resident Ca2+-binding protein with multiple EF-hand motifs and a carboxyl-terminal HDEL sequence. J Biol Chem 1993; 268:699-705.
Weis K, Griffiths G, Lamond Al. The endoplasmic reticulum calcium-binding protein of 55 kDa is a novel EF-hand protein retained in the endoplasmic reticulum by a carboxyl-terminal His-Asp-Glu-Leu motif. J Biol Chem 1994; 269:19142-19150.
Pearson RB, Kemp BE. Protein kinase phosphorylation site sequences and consensus specificity motifs: tabulations. In: Hunter T. Sefton BM (eds.), Methods in Enzymology, Part A, Vol. 200. San Diego: Academic Press Inc.; 1991: 62-81.
Robbi M, Beaufay H. The COOH terminus of several liver carboxylesterases targets these enzymes to the lumen of the endoplasmic reticulum. J Biol Chem 1991; 266:20498-20503.
Pelham HRB. The dynamic organisation of the secretory pathway. Cell Struct Funct 1996; 21:413-419.
Munro S, Pelham HRB. A C-terminal signal prevents secretion of luminal ER proteins. Cell 1987; 48:899-907.
Tang BL, Wong SH, Qi XL, Low SH, Hong W. Molecular cloning, characterization, subcellular localization and dynamics of p23, the mammalian KDEL receptor. J Cell Biol 1993; 120:325-338.
Yoshimori T, Semba T, Takemoto H, Akagi S, Yamamoto A, Tashiro Y. Protein disulfide-isomerase in rat exocrine pancreatic cells is exported from the endoplasmic reticulum despite possessing the retention signal. J Biol Chem 1990; 265:15984-15990.
Takemoto H, Yoshimori T, Yamamoto A, Miyata Y, Yahara I, Inoue K, Tashiro Y. Heavy chain binding protein (BiP/GRP78) and endoplasmin are exported from the endoplasmic reticulum in rat exocrine pancreatic cells, similar to protein disulfide-isomerase. Arch Biochem Biophys 1992; 296:129-136.
Alexandropoulos K, Cheng GH, Baltimore D. Proline-rich sequences that bind to Src homology 3 domains with individual specificities. Proc Natl Acad Sci USA 1995; 92:3110-3114.
Ophoff RA, Terwindt GM, Vergouwe MN, Van Eijk R, Mohrenweiser H. Litt M, Hofker MH, Haan J, Ferrari MD, Frants RR. A 3-Mb region for the familial hemiplegic migraine locus on 19p13.1-p13.2: exclusion of PRKCSH as a candidate gene. Eur J Hum Genet 1996; 4:312-328.
Nykjaer A, Dragun D, Walther D, Vorum H, Jacobsen C, Herz J, Melsen F, Christensen El, Willnow E. An endocytotic pathway essential for renal uptake and activation of the steroid 25-(OH) vitamin D3. Cell 1999; 96:507-515.
Hammond C, Braakman I, Helenius A. Role of N-linked oligosaccharide recognition, glucose trimming, and calncxin in glycoprotein folding and quality control. Proc Natl Acad Sci USA 1994; 91:913-917.
Saxena S, Shailubhai K, Dong-Yu B, Vijay IK. Purification and characterization of glucosidase II involved in N-linked glycoprotein processing in bovine mammary gland. Biochem J 1987; 247:563-570.
Burns DM, Touster O. Purification and characterization of glucosidase II, and endoplasmic reticulum hydrolase involved in glycoprotein biosynthesis. J Biol Chem 1982; 257:9991-10000.
Strous GJ, Van Kerkhof P, Brok R, Roth J, Brada D. Glucosidase II, a protein of the endoplasmic reticulum with high mannose oligosaccharide chains and a rapid turnover. J Biol Chem 1987; 262:3620-3625.
Lucocq JM, Brada D, Roth J. Immunolocalization of the oligosaccharide trimming enzyme glucosidase II. J Cell Biol 1986; 102:2137-2146.
Brada D, Kerjaschki D, Roth J. Cell type-specific post-Golgi apparatus localization of a "resident" endoplasmic reticulum glycoprotein, glucosidase II. J Cell Biol 1990; 110:309-318.
