The preparation of calmodulins from barley (Hordeum sp.) and basidiomycete fungi.
1. Calmodulin-like proteins were purified from the fruiting bodies of higher (basidiomycete) fungi and barley (Hordeum sp.) shoots. 2. These calmodulins have electrophoretic mobilities on 10% (w/v) polyacrylamide gels at pH 8.3 in the presence of 6 M-urea and at pH 8.3 in the presence of 0.1% sodium dodecyl sulphate similar to that of bovine brain calmodulin. They interacted with rabbit skeletal-muscle troponin I in the presence of Ca2+. 3. Barley and fungal calmodulins activated myosin light-chain kinase and phosphodiesterase in the presence of Ca2+, although the amounts needed were at least an order of magnitude greater than is required to produce the same effect with mammalian calmodulin. 4. Amino acid analyses indicated a number of differences from the mammalian protein, most notably the absence of trimethyl-lysine. 5. By using 125I-labelled calmodulin, a small amount of calmodulin-binding protein was detected in homogenates of barley and fungi. 6. No protein corresponding to calmodulin could be found in Escherichia coli or yeast, although a relatively high concentration of a protein that bound calmodulin was detected in E. coli by this technique.
D015105:3',5'-Cyclic-AMP Phosphodiesterases / Q000378:metabolism; D000596:Amino Acids / Q000032:analysis; D001487:Basidiomycota / Q000032:analysis*; D002135:Calcium-Binding Proteins / Q000302:isolation & purification*; D002147:Calmodulin / Q000302:isolation & purification* / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation / Q000187:drug effects; D001467:Hordeum / Q000032:analysis; D009219:Myosin-Light-Chain Kinase; D010944:Plants / Q000032:analysis*; D011494:Protein Kinases / Q000378:metabolism; D014336:Troponin / Q000378:metabolism
D000596:Amino Acids; D002135:Calcium-Binding Proteins; D002147:Calmodulin; D014336:Troponin; D011494:Protein Kinases; D009219:Myosin-Light-Chain Kinase; D015105:3',5'-Cyclic-AMP Phosphodiesterases