2009年7月
Biochemical Interaction of an Actin-Capping Protein, CapZ, with NAP-22
JOURNAL OF NEUROSCIENCE RESEARCH
- ,
- ,
- ,
- 巻
- 87
- 号
- 9
- 開始ページ
- 1980
- 終了ページ
- 1985
- 記述言語
- 英語
- 掲載種別
- 研究論文(学術雑誌)
- DOI
- 10.1002/jnr.22040
- 出版者・発行元
- WILEY-LISS
NAP-22 is a neuronal protein localized in the presynaptic membrane and synaptic vesicles and recovered in a Triton-insoluble low-density microdomain fraction after biochemical fractionation of the synaptic plasma membrane. NAP-22 organizes membrane microdomains through binding to membrane lipids such as cholesterol, phosphatidylethanolamine, and phosphatidylinositol 4,5-bisphosphate. In this study, NAP-22-binding proteins were screened through the pull-down assay using brain-derived NAP-22 bound to Sepharose 4B. An actin-capping protein, CapZ, was identified in the precipitate through mass spectrometry and Western blotting. CapZ was then expressed in E coli and the purified protein-bound NAP-22 directly. Because bacterially expressed NAP-22 bound CapZ, it was determined that the N-terminal myristoyl moiety of NAP-22 is not necessary for the binding. The binding of NAP-22 showed no effect on the actin nucleation activity of CapZ measured with centrifugation and viscometric assays. Hence, the CapZ-NAP-22 complex could work as the nucleation site of actin polymerization or as the actin filament-anchoring site on the membrane microdomain. (C) 2009 Wiley-Liss, Inc.
- リンク情報
- ID情報
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- DOI : 10.1002/jnr.22040
- ISSN : 0360-4012
- PubMed ID : 19267422
- Web of Science ID : WOS:000266468300004