Document Detail


G protein gamma subunits coimmunoprecipitated with antibodies against alpha subunits: identification of major isoforms in cultured cells by silver stain and immunoblotting with conventional transfer procedure.
MedLine Citation:
PMID:  9792930     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The betagamma subunits of G proteins were coimmunoprecipitated with antibodies against various alpha subunits, and analyzed by silver stain and immunoblotting with conventional transfer procedure and membrane-blocking buffer containing 2% BSA. Multiple isoforms of gamma were coimmunoprecipitated with no significant difference in form or ratio among the antibodies against alpha subunits used, suggesting antibodies against any alpha subunit could coimmunoprecipitate all forms of gamma. Therefore, this method was applicable to analyze gamma subunits in various cells, especially to clarify what forms of gamma subunits are major components. The major isoforms were: gamma5 in C6, NG108-15, HeLa, HEK293, and F9 cells; gamma12 in Swiss 3T3 and BRL-3A cells; and gamma3 in PC12 cells. In addition to most gamma subunits identified, unidentified gamma subunits were present in PC12, NG108-15, and BRL-3A cells. Furthermore, the method was applied to examine changes of isoforms of gamma during differentiation of HL-60 cells. Undifferentiated cells mainly contained gamma5, but retinoic acid treatment of cells replaced most gamma5 with gamma2. Thus, this method is useful to determine the major isoforms which seem to be the more important in cells.
Authors:
H Ueda; R Morishita; R Katoh-Semba; K Kato; T Asano
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of biochemistry     Volume:  124     ISSN:  0021-924X     ISO Abbreviation:  J. Biochem.     Publication Date:  1998 Nov 
Date Detail:
Created Date:  1999-02-11     Completed Date:  1999-02-11     Revised Date:  2007-12-19    
Medline Journal Info:
Nlm Unique ID:  0376600     Medline TA:  J Biochem     Country:  JAPAN    
Other Details:
Languages:  eng     Pagination:  1033-7     Citation Subset:  IM    
Affiliation:
Department of Biochemistry Institute for Developmental Research, Aichi Human Service Center, Kasugai, Aichi, 480-0392, Japan.
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Animals
Antibodies / immunology
Blotting, Western
Cell Line
GTP-Binding Proteins / chemistry,  immunology,  metabolism*
Humans
Precipitin Tests
Silver Staining
Chemical
Reg. No./Substance:
0/Antibodies; EC 3.6.1.-/GTP-Binding Proteins

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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