Bioquimica y patologia clinica : ByPC : revista de la Asociacion Bioquimica Argentina最新文献

筛选

英文中文

Purification and molecular characterization of NP185, a neuronal-specific and synapse-enriched clathrin assembly polypeptide. 神经元特异性和富含突触的网格蛋白组装多肽NP185的纯化和分子特性。

Bioquimica y patologia clinica : ByPC : revista de la Asociacion Bioquimica Argentina Pub Date : 1998-01-01

Shengwen Li, Michael Lisanti, Saul Puszkin

{"title":"Purification and molecular characterization of NP185, a neuronal-specific and synapse-enriched clathrin assembly polypeptide.","authors":"Shengwen Li, Michael Lisanti, Saul Puszkin","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>NP185, a neuronal-specific protein of 185 kDa, was first discovered when we prepared monoclonal anti-bodies (mAbs) against bovine brain clathrin coated vesicles. Two mAbs, 8G8 and 6G7, permitted us to characterize this protein both biochemically and in development (NP185 is expressed in a NGF-dependent manner in PC12 cells). The expression of NP185 coincides with synaptogenesis. In this work, we have further characterized this protein as follows: Microsequence analysis of immuno-purified native NP185 from bovine brain yielded five peptides that corresponded exactly to the known sequences of murine F1-20 and rat AP180 (renamed AP3); ii) Using an established assay, we show that purified recombinant NP185/AP3 can facilitate clathrin cages assembly; iii) Using deletion mutagenesis, we mapped the epitopes of two distinct mAbs directed against bovine NP185 to a 60 amino acid residue region of the murine recombinant NP185/AP3; iv) Recombinant NP185/AP3 can be phosphorylated by purified casein kinase II in vitro; and v) Recombinant NP185/AP3 directly binds to purified brain tubulin. Since NP185/AP3 binds to tubulin and stimulates the clathrin assembly, it may be involved in the regulation of the transport of clathrin-coated vesicles. Casein kinase II, an enzyme known to be present in clathrin-coated vesicles, may play a role in the regulation of NP185/AP3 for the promotion of clathrin assembly.</p>","PeriodicalId":92758,"journal":{"name":"Bioquimica y patologia clinica : ByPC : revista de la Asociacion Bioquimica Argentina","volume":"62 1","pages":"5-17"},"PeriodicalIF":0.0,"publicationDate":"1998-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6688760/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147517533","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0