دورية أكاديمية
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.
| العنوان: | 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. |
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| المؤلفون: | Brûlé, Sophie, Rabahi, Flora, Faure, Robert, Beckers, Jean-François, Silversides, David W., Lussier, Jacques G. |
| المصدر: | Biology of Reproduction, 62 (3), 642 - 654 (2000-03) |
| بيانات النشر: | Society for the Study of Reproduction, 2000. |
| سنة النشر: | 2000 |
| مصطلحات موضوعية: | Calcium-Binding Proteins, DNA, Complementary, Intracellular Signaling Peptides and Proteins, Membrane Proteins, Phosphoproteins, Prkcsh protein, mouse, Prkcsh protein, rat, Recombinant Proteins, VASAP-60 protein, Bos taurus, Myristoylated Alanine-Rich C Kinase Substrate, Glucosidases, PRKCSH protein, human, beta-Glucosidase, Amino Acid Sequence, Animals, Antibody Specificity, Base Sequence, Cattle, Corpus Luteum/metabolism, Female, Gene Expression Regulation, Glycosylation, Granulosa Cells/metabolism, Humans, Intracellular Membranes/metabolism, Membrane Proteins/genetics, Membrane Proteins/immunology, Membrane Proteins/metabolism, Mice, Molecular Sequence Data, Phosphoproteins/metabolism, Rats, Recombinant Proteins/genetics, Recombinant Proteins/metabolism, Sequence Analysis, DNA, Sequence Homology, Amino Acid, Vacuoles/metabolism, beta-Glucosidase/metabolism, Reproductive Medicine, Cell Biology, General Medicine, Life sciences, Veterinary medicine & animal health, Sciences du vivant, Médecine vétérinaire & santé animale |
| الوصف: | 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. |
| نوع الوثيقة: | journal article http://purl.org/coar/resource_type/c_6501Test article peer reviewed |
| اللغة: | English |
| العلاقة: | http://academic.oup.com/biolreprod/article-pdf/62/3/642/10594715/biolreprod0642.pdfTest; urn:issn:0006-3363; urn:issn:1529-7268 |
| DOI: | 10.1095/biolreprod62.3.642 |
| الوصول الحر: | https://orbi.uliege.be/handle/2268/298359Test |
| حقوق: | restricted access http://purl.org/coar/access_right/c_16ecTest info:eu-repo/semantics/restrictedAccess |
| رقم الانضمام: | edsorb.298359 |
| قاعدة البيانات: | ORBi |
| DOI: | 10.1095/biolreprod62.3.642 |
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