Suramin and the suramin analogue NF307 discriminate among calmodulin-binding sites.
Calmodulin-binding sites on target proteins show considerable variation in primary sequence; hence compounds that block the access of calmodulin to these binding sites may be more selective than compounds that inactivate calmodulin. Suramin and its analogue NF307 inhibit the interaction of calmodulin with the ryanodine receptor. We have investigated whether inhibition of calmodulin binding to target proteins is a general property of these compounds. Suramin inhibited binding of [(125)I]calmodulin to porcine brain membranes and to sarcoplasmic reticulum from skeletal muscle (IC(50)=4.9+/-1.2 microM and 19.9+/-1.8 microM, respectively) and blocked the cross-linking of [(125)I]calmodulin to some, but not all, target proteins in brain membranes by [(125)I]calmodulin. Four calmodulin-binding proteins were purified [ryanodine receptor-1 (RyR1) from rabbit skeletal muscle, neuronal NO synthase (nNOS) from Sf9 cells, G-protein betagamma dimers (Gbetagamma) from porcine brain and a glutathione S-transferase-fusion protein comprising the C-terminal calmodulin-binding domain of the metabotropic glutamate receptor 7A (GST-CmGluR7A) from bacterial lysates]. Three of the proteins employed (Gbetagamma, GST-CmGluR7A and RyR1) display a comparable affinity for calmodulin (in the range of 50-70 nM). Nevertheless, suramin and NF307 only blocked the binding of Gbetagamma and RyR1 to calmodulin-Sepharose. In contrast, the association of GST-CmGluR7A and nNOS was not impaired, whereas excess calmodulin uniformly displaced all proteins from the matrix. Thus suramin and NF307 are prototypes of a new class of calmodulin antagonists that do not interact directly with calmodulin but with calmodulin-recognition sites. In addition, these compounds discriminate among calmodulin-binding motifs.
Klinger
M
M
Institute of Pharmacology, University of Vienna, Währinger Strasse 13a, A-1090 Vienna, Austria.
Bofill-Cardona
E
E
Mayer
B
B
Nanoff
C
C
Freissmuth
M
M
Hohenegger
M
M
D000494:Allosteric Regulation; D020816:Amino Acid Motifs; D000818:Animals; D001665:Binding Sites; D001921:Brain / Q000187:drug effects* / Q000378:metabolism; D002147:Calmodulin / Q000187:drug effects / Q000378:metabolism*; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002846:Chromatography, Affinity; D003432:Cross-Linking Reagents / Q000378:metabolism; D066298:In Vitro Techniques; D007457:Iodine Radioisotopes; D018482:Muscle, Skeletal / Q000187:drug effects / Q000378:metabolism; D009282:Naphthalenesulfonates / Q000494:pharmacology*; D009842:Oligopeptides / Q000494:pharmacology*; D011817:Rabbits; D012519:Sarcoplasmic Reticulum / Q000187:drug effects* / Q000378:metabolism; D013498:Suramin / Q000494:pharmacology*; D013552:Swine
D002147:Calmodulin; D003432:Cross-Linking Reagents; D007457:Iodine Radioisotopes; C117862:NF 307; D009282:Naphthalenesulfonates; D009842:Oligopeptides; D013498:Suramin