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PMID: 228291 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Photoaffinity labeling of cyclic-AMP- and AMP-binding proteins differentiating Dictyostelium discoideum cells.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 76 ·No. 9 ·1979-09-00 ·Pages 4250-4

Wallace LJ, Frazier WA

Abstract

Cyclic-AMP-binding proteins play important roles during the differentiation of the cellular slime mold Dictyostelium discoideum. The photoaffinity reagent 8-N3-cyclic [32P]AMP has been used to label developmentally regulated cyclic-AMP-binding proteins of intact cells, membranes, and cytoplasm. 8-N3-Cyclic AMP is a chemoattractant for differentiated D. discoideum cells and is a substrate for the membrane phosphodiesterase (mPDE). When mPDE is inhibited, the only specifically labeled protein on intact cells has a molecular weight of 40,000 on sodium dodecyl sulfate gels. The developmental time course of appearance of this protein and its high specificity for cyclic AMP identify it as the cell surface chemotactic receptor for cyclic AMP. The concentration dependence of labeling of this protein is consistent with the measured chemotactic potency of 8-N3-cyclic AMP, which is about 1/100th that of cyclic AMP. Three developmentally regulated proteins (Mr 26,000, 33,000, and 36,000) of the soluble fraction (cytoplasm) are labeled by the photoaffinity reagent and are specific for cyclic AMP. By analogy with other systems, these may be regulatory subunits of protein kinases. The mPDE of ghosts or plasma membrane fractions converts the reagent to 8-N3-[32P]AMP, which specifically photoaffinity labels a protein of Mr 42,000 associated with the cytoplasmic face of the plasma membrane.

MeSH Terms
Affinity Labels Cell Differentiation Cell Membrane/metabolism Chemotaxis Cyclic AMP/analogs & derivatives,metabolism Dictyostelium/cytology,metabolism Dose-Response Relationship, Drug Membrane Proteins/metabolism Molecular Weight Receptors, Cyclic AMP/metabolism
Chemicals
Affinity Labels Membrane Proteins Receptors, Cyclic AMP Cyclic AMP
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Wallace L J
Frazier W A
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38 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.76.9.4250
Published
1979-09-00
Pages
4250-4
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC411550
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.76.9.4250 PMC Full Text View journal details
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