期刊
JOURNAL OF MOLECULAR BIOLOGY
卷 419, 期 5, 页码 359-368出版社
ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD
DOI: 10.1016/j.jmb.2012.03.015
关键词
actin filament; electron tomography; polarity of the actin filament; image analysis; cytoskeleton
资金
- Ministry of Education, Culture, Sports, Science and Technology of the Japanese Government
- Daiko Research Foundation
- Austrian Science Fund [FWF 1516-B09, FWF P21292-B09]
- Austrian Science Fund (FWF) [P 21292] Funding Source: researchfish
- Grants-in-Aid for Scientific Research [20227008, 22770145] Funding Source: KAKEN
- Austrian Science Fund (FWF) [P21292] Funding Source: Austrian Science Fund (FWF)
Actin filaments are polar structures that exhibit a fast growing plus end and a slow growing minus end. According to their organization in cells, in parallel or antiparallel arrays, they can serve, respectively, in protrusions or in contractions. The determination of actin filament polarity in subcellular compartments is therefore required to establish their local function. Myosin binding has previously been the sole method of polarity determination. Here, we report the first direct determination of actin filament polarity in the cell without myosin binding. Negatively stained cytoskeletons of lamellipodia were analyzed by adapting electron tomography and a single particle analysis for filamentous complexes. The results of the stained cytoskeletons confirmed that all actin filament ends facing the cell membrane were the barbed ends. In general, this approach should be applicable to the analysis of actin polarity in tomograms of the actin cytoskeleton. (C) 2012 Elsevier Ltd. All rights reserved.
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