Characteristics of rabbit muscle adenylate kinase inhibition by sulfur and recovery by dithiothreitol.
Structure-function relationships of rabbit muscle adenylate kinase (RMAK) were studied by examining the characteristics of inhibitions by hydrophobic inhibitors and reactivations by sulfhydryl reagents. RMAK is inhibited by 1-butanol,N-ethylmaleimide (NEM) and elemental sulfur (S8) with increasing effectiveness in the order of increasing hydrophobicity. Characteristics of these hydrophobic inhibitors are compared with inhibitors forming covalent bonds or reversible complexes. A mechanism is proposed for hydrophobic inhibitors of RMAK that involves conformational changes promoted by interacting with hydrophobic regions. The reversal of RMAK inhibition by sulfhydryl compounds involves a conformational change that exposes hydrophobic regions and the inhibitor to water. Circular dichroism (CD) data show changes in the secondary structures of RMAK, indicating that the inhibitors and the sulfhydryl compounds promote conformational changes. The results of these studies show that the activity of a small enzyme can be controlled in a manner analogous to the allosteric control of larger enzymes.
Russell
P J
PJ
Department of Biology, University of California, San Diego, La Jolla 92093, USA.
Williams
A
A
Avila
D
D
Chinn
E
E
Taulane
J P
JP
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000263:Adenylate Kinase / Q000037:antagonists & inhibitors* / Q000737:chemistry; D000438:Alcohols / Q000494:pharmacology; D000818:Animals; D001665:Binding Sites; D002942:Circular Dichroism; D004228:Dithionitrobenzoic Acid / Q000494:pharmacology; D004229:Dithiothreitol / Q000494:pharmacology*; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005033:Ethylmaleimide / Q000494:pharmacology; D006902:Hydroxymercuribenzoates / Q000494:pharmacology; D007700:Kinetics; D018482:Muscle, Skeletal / Q000201:enzymology*; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D011817:Rabbits; D012835:Silver Nitrate / Q000494:pharmacology; D013329:Structure-Activity Relationship; D013438:Sulfhydryl Compounds / Q000494:pharmacology; D013455:Sulfur / Q000494:pharmacology*
D000438:Alcohols; D004791:Enzyme Inhibitors; D006902:Hydroxymercuribenzoates; D013438:Sulfhydryl Compounds; C030601:4-hydroxymercuribenzoate; D013455:Sulfur; D000255:Adenosine Triphosphate; D012835:Silver Nitrate; D004228:Dithionitrobenzoic Acid; D000263:Adenylate Kinase; D005033:Ethylmaleimide; D004229:Dithiothreitol