Document Detail

In vitro modulation of filament bundling in F-actin and keratins by annexin II and calcium.
MedLine Citation:
PMID:  7520812     Owner:  NLM     Status:  MEDLINE    
In our preliminary subcellular localization experiment we demonstrated that annexin II co-localized with submembranous actin in subpopulations of both cultured fibroblasts and keratinocytes. To investigate the physical interaction between annexin II and actin at the cell periphery, in vitro reconstitution experiments were carried out with keratins used as a control. Annexin II, isolated by immunoaffinity column chromatography, was found to exist as globular structures measuring 10 to 25 nm in diameter by rotary shadowing, similar to a previous report. We believe that these structures represent its polymeric forms. By negative staining, monomeric annexin II was detectable as tapered rods, measuring 6 nm in length and 1 to 2 nm in diameter. When annexin II was mixed with actin in 3 mM piperazine-N, N-bis-2-ethanesulfonic acid (PIPES) buffer with 10 mM NaCl2, 2 mM MgCl2 and 0.1 mM CaCl2, thick twisting actin bundles formed, confirming previous reports. This bundling was much reduced when calcium was removed. In the presence of 5 mM ethylenediamine tetra-acetic acid (EDTA) in 5 mM tris, pH 7.2, keratins were found to form a network of filaments, which began to disassemble when the chelator was removed and became fragmented when 0.1 mM CaCl2 was added. Keratins under the same conditions did not fragment when annexin II was present. These results suggest that annexin II, in conjunction with Ca2+, may be involved in a flexible system accommodating changes in the membrane cytoskeletal framework at the cell periphery in keratinocytes.
A S Ma; M E Bystol; A Tranvan
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  In vitro cellular & developmental biology. Animal     Volume:  30A     ISSN:  1071-2690     ISO Abbreviation:  In Vitro Cell. Dev. Biol. Anim.     Publication Date:  1994 May 
Date Detail:
Created Date:  1994-09-27     Completed Date:  1994-09-27     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  9418515     Medline TA:  In Vitro Cell Dev Biol Anim     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  329-35     Citation Subset:  IM    
Department of Medicine, University of Chicago, Illinois 60637.
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MeSH Terms
3T3 Cells
Actins / metabolism*
Annexin A2 / isolation & purification,  metabolism*,  ultrastructure
Calcium / metabolism*
Cells, Cultured
Chromatography, Affinity
Electrophoresis, Polyacrylamide Gel
Keratinocytes / metabolism
Keratins / metabolism*
Grant Support
Reg. No./Substance:
0/Actins; 0/Annexin A2; 68238-35-7/Keratins; 7440-70-2/Calcium

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