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Detection of multiple forms of G iα in HL60 cells
Author(s) -
Murphy Philip M.,
Eide Brock,
Goldsmith Paul,
Brann Mark,
Gierschik Peter,
Spiegel Allen,
Malech Harry L.
Publication year - 1987
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(87)80356-3
Subject(s) - complementary dna , biology , blot , microbiology and biotechnology , northern blot , protein subunit , southern blot , adenylate kinase , g protein , western blot , oligonucleotide , biochemistry , gene , enzyme , receptor
Comparison of cDNA sequences from multiple sources predicts a genus of highly homologous but structurally distinct G protein α‐subunits, designated as G iα , that may include the α‐subunit of the functionally defined adenylate cyclase inhibitory G protein. Using specific oligonucleotide probes on Northern blots, we show that G iα−2 and G iα−3 , but not G iα−1 , are expressed in HL60 cells. Antisera raised against synthetic peptides from regions predicted to be conserved (AS6) and divergent (LE3) among G iα subtypes bind to a 40 kDa protein in Western blots of HL60 membranes. AS6 identifies an additional protein at 41 kDa. Thus, Northern blot and immunoblot results show that at least two G iα subtypes, a 40 kDa protein likely to correspond to G iα−2 and a 41 kDa protein possibly representing G iα−3 , may be expressed in a single cell type.