Summary
Adenylate cyclase from rat hippocampus was separated by electrophoresis in polyacryl amide microgels and stained for enzymatic activity using a new histochemical procedure. This method involves the use of AMP-PNP, aminophylline, dithiotreitole, and Sr2+ as “primary” capture ions, thus fulfilling all the demands for a really specific histochemical incubation medium for the enzyme. The incubation of the gels with this medium resulted in the inhibition of other enzymes, which are capable of splitting AMP-PNP (ATP: pyrophosphatase, alkaline phosphatase), whereas adenylate cyclase remained highly active under these conditions. The enzyme was found to be present in two forms in the gels. Both protein bands were stimulated by the addition of various biogenic amines to the incubation medium. One protein band was fully GMP-PNP dependent in its activity. It is reasonable to suppose that these forms are either differently high aggregated molecules of the enzyme or enzyme molecules bound to their regulatory sites.
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Poeggel, G., Bernstein, H.G., Luppa, H. et al. Specific demonstration of rat brain adenylate cyclase in polyacryl amide microgels by a new histochemical procedure. Histochemistry 73, 305–309 (1981). https://doi.org/10.1007/BF00493028
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DOI: https://doi.org/10.1007/BF00493028