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PMID: 2668303 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Association of type 3 protein kinase C with focal contacts in rat embryo fibroblasts.

The Journal of cell biology ·Vol. 109 ·No. 2 ·1989-08-00 ·Pages 697-704

Jaken S, Leach K, Klauck T

Abstract

We have used immunocytofluorescence techniques to determine the subcellular distribution of the Ca2+, phospholipid-dependent protein kinase, protein kinase C (PKC). Using monoclonal antibodies that are specific for Type 3 (alpha) PKC, we have determined that there are least two pools of PKC in normal rat embryo fibroblasts (REF52 cells): diffuse cytoplasmic and fiber-associated. Extraction with chelators and detergent before fixing and staining removes the cytoplasmic PKC. The fiber-associated staining remains in these cytoskeleton preparations. The cytoskeleton Type 3 PKC staining closely resembles that of the focal contact protein vinculin and colocalizes with another focal contact protein, talin. Cytochalasin, but not colchicine, coordinately disrupts the staining pattern of vinculin and PKC. Activation of PKC by treatment with phorbol esters causes depolymerization of microfilaments and reorganization of vinculin staining. We propose that Type 3 PKC is a modulatory component of the focal contact and has a primary role in regulation of the association of microfilament bundles with the plasma membrane.

MeSH Terms
Actin Cytoskeleton/drug effects,metabolism,ultrastructure Animals Antibodies, Monoclonal/immunology Cell Membrane/drug effects,metabolism,ultrastructure Cells, Cultured Colchicine/pharmacology Cytochalasin B/pharmacology Cytoskeletal Proteins/metabolism Cytoskeleton/drug effects,metabolism,ultrastructure Embryo, Mammalian/cytology,metabolism Fibroblasts/cytology,metabolism,ultrastructure Fluorescent Antibody Technique Immunohistochemistry Isoenzymes/immunology,metabolism Phorbol 12,13-Dibutyrate/pharmacology Phosphorylation Protein Kinase C/immunology,metabolism Rats
Chemicals
Antibodies, Monoclonal Cytoskeletal Proteins Isoenzymes Phorbol 12,13-Dibutyrate Cytochalasin B Protein Kinase C Colchicine
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Jaken S
W. Alton Jones Cell Science Center, Inc., Lake Placid, New York 12946.
Leach K
Klauck T
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.109.2.697
Published
1989-08-00
Pages
697-704
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2115725
Subset
IM
Grants
NCI NIH HHS · CA48137 · United States
External Links
PubMed source DOI: 10.1083/jcb.109.2.697 PMC Full Text View journal details
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