Identification of Ca2+-dependent calmodulin-binding proteins in rat spermatogenic cells as complexes of the heat-shock proteins
| Autor: | Megumi Moriya, Michio Yazawa, Hajime J. Yuasa, Norio Suzuki, Masanori Ochiai |
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| Rok vydání: | 2004 |
| Předmět: |
Male
Protein family Blotting Western Molecular Sequence Data Fluorescent Antibody Technique Spermatocytes Heat shock protein Protein A/G HSPA2 Genetics Animals Amino Acid Sequence Spermatogenesis Peptide sequence Heat-Shock Proteins biology Cell Biology Immunohistochemistry Spermatids Calmodulin-binding proteins Rats Cell biology Hsp70 Biochemistry Cytoplasm biology.protein Calcium Calmodulin-Binding Proteins Developmental Biology |
| Zdroj: | Molecular Reproduction and Development. 69:316-324 |
| ISSN: | 1098-2795 1040-452X |
| DOI: | 10.1002/mrd.20134 |
| Popis: | Ca2+-calmodulin (CaM)-binding proteins in rat testes were characterized by assays for CaM-binding activity using the CaM-overlay method on transblots of electrophoresed gels and purification by gel-filtration, ion exchange, and adsorption chromatographies. A major CaM-binding protein complex (CaMBP) was identified and found to be comprised of three proteins with molecular masses 110, 100, and 70 kDa. Amino acid sequence analyses of lysylendopeptidase digests from these proteins indicated that all of the constituents of CaMBP are very similar to the members of the heat-shock protein family, i.e., the 110-kDa protein is similar to the APG-2/94 kDa rat ischemia-responsive protein, the 100-kDa protein is similar to the rat counterpart of the mouse APG-1/94 kDa osmotic stress protein, and the 70-kDa protein is similar to the rat testis-specific major heat-shock protein (HSP70). Immunohistochemistry using anti-CaMBP and anti-CaM antibodies demonstrated that CaMBP was co-localized with CaM in the cytoplasm of pachytene spermatocytes and nuclei of round spermatids. In addition, CaMBP, but not CaM, was localized at a high level in the residual bodies of elongated spermatids. The possible relevance of CaMBP to regulation of cell cycle progression and spermatogenesis is discussed in this paper. Mol. Reprod. Dev. 69: 316–324, 2004. © 2004 Wiley-Liss, Inc. |
| Databáze: | OpenAIRE |
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