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Adenylate cyclase and guanylate cyclase in the excitable ciliary membrane from Paramecium: Separation and regulation
Author(s) -
Klumpp Susanne,
Gierlich Doris,
Schultz Joachim E.
Publication year - 1984
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(84)80466-4
Subject(s) - cyclase , adenylate kinase , gucy1a3 , egta , paramecium , enzyme , gucy1b3 , chemistry , calmodulin , biochemistry , centrifugation , protein subunit , membrane , differential centrifugation , chromatography , guanylate cyclase 2c , calcium , organic chemistry , gene
Particulate adenylate cyclase (AC) and guanylate cyclase (GC) activities localized in the ciliary membrane from Paramecium were solubilized by a two‐step procedure using the detergents Brij 56 and Lubrol PX. The enzymes remained in the supernatant after a 100 000 × g centrifugation. Upon gel chromatography, AC and GC were almost completely separated proving that each enzyme is a distinct molecular entity. Solubilization of GC was achieved with the calmodulin subunit remaining firmly attached to the catalytic part. Antibodies against calmodulin inhibited the enzyme as did La 3+ and EGTA. AC activity appeared to be regulated specifically by K + , enzyme activity being enhanced up to 100% by 15 mM K + . Na + and Li + were inactive.