Regulation of skeletal-muscle AMP deaminase: involvement of histidine residues in the pH-dependent inhibition of the rabbit enzyme by ATP.
Reaction of rabbit skeletal-muscle AMP deaminase with a low molar excess of diethyl pyrocarbonate results in conversion of the enzyme into a species with one or two carbethoxylated histidine residues per subunit that retains sensitivity to ATP at pH 7.1 but, unlike the native enzyme, it is not sensitive to regulation by ATP at pH 6.5. This effect mimics that exerted on the enzyme by limited proteolysis with trypsin, which removes the 95-residue N-terminal region from the 80 kDa enzyme subunit. These observations suggest involvement of some histidine residues localized in the region HHEMQAHILH (residues 51-60) in the regulatory mechanism which stabilizes the binding of ATP to its inhibitory site at acidic pH. Carbethoxylation of two histidine residues per subunit abolishes the inhibition by ATP of the proteolysed enzyme at pH 7.1, suggesting the obligatory participation of a second class of histidine residues, localized in the 70 kDa subunit core, in the mechanism of the pH-dependent inhibition of the enzyme by ATP. At a slightly acidic pH, these histidine residues would be positively charged, resulting in a desensitized form of the enzyme similar to that obtained with the carbethoxylation reaction.
Ranieri-Raggi
M
M
Istituto di Chimica Biologica dell'Università di Pisa, Italy.
Ronca
F
F
Sabbatini
A
A
Raggi
A
A
D000659:AMP Deaminase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000494:pharmacology*; D000494:Allosteric Regulation; D000595:Amino Acid Sequence; D000818:Animals; D004047:Diethyl Pyrocarbonate / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000494:pharmacology; D006639:Histidine / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000201:enzymology*; D011817:Rabbits; D014357:Trypsin / Q000494:pharmacology
D006639:Histidine; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D014357:Trypsin; D000659:AMP Deaminase; D004047:Diethyl Pyrocarbonate