Premium
Polymorphism of soluble rat brain guanylate cyclase is produced during isoelectric focusing
Author(s) -
Basset Paul,
Zwiller Jean,
Revel MarieOdile,
Vincendon Guy
Publication year - 1983
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150040606
Subject(s) - isoelectric focusing , chemistry , enzyme , biochemistry , guanylate cyclase , chromatography , gel electrophoresis , isoelectric point , polyacrylamide gel electrophoresis
Soluble guanylate cyclase purified from rat brain exhibits about 30 different components between pH 5.3 and 6.5 on polyacrylamide gel isoelectric focusing (IEF) Specific immunodetection after IEF of guanylate cyclase from fresh rat brain supernatant allows the detection of a complex profile, indicating that the complex pattern observed with purified enzyme is not related to sample aging. In contrast, purified enzyme gives only one major band on electrophoresis in presence or absence of sodium dodecyl sulfate and a single line of immunoprecipitation was obtained on crossed immunoelectrofocusing. The IEF pattern exhibited by guanylate cyclase is dependent on the sample concentration, the pH range of sample application, the carrier ampholytes used and on the presence of urea in the gel. Fractionation tests, performed with partially purified enzyme or with purified enzyme, indicate that this complex pattern is produced during IEF and does not correspond to an inherent heterogeneity of soluble rat brain guanylate cyclase. The distribution of carrier ampholytes used to form the pH gradient and that of guanylate cyclase bands was found to be very similar at the steady‐state of IEF, suggesting that interaction between carrier ampholytes and rat brain soluble guanylate cyclase can be involved in the generation of the complex IEF pattern.