AMP-deaminase from normal and cirrhotic human liver: a comparative study.
AMP-deaminase (EC 3.5.4.6) is an enzyme of nucleotide breakdown involved in regulation of adenine nucleotide pool in mammalian cells. Reaction catalysed by AMP-deaminase constitutes a rate-limiting step in adenine nucleotide catabolism in liver. In this study kinetic and regulatory properties of AMP-deaminase purified from normal and cirrhotic human liver were investigated. In comparison to AMP-deaminase extracted from the normal human liver, AMP-deaminase extracted from the cirrhotic liver was less sensitive towards substrate analogues, and only a very limited response towards pH and adenylate energy charge changes tested for enzyme isolated from this tissue source had been observed. At physiological pH 7.0, in the absence and in the presence of important allosteric effectors (ATP, ADP, GTP and orthophosphate), AMP-deaminases from the two sources studied manifested different regulatory profiles, with half-saturation constant (S0.5) values being distinctly higher for the enzyme extracted from the pathological organ. In contrast to AMP-deaminase isolated from the normal, healthy liver, where presence of relatively large (68 kDa) protein fragment was also detected, only smaller protein fragments were identified, while SDS-PAG electrophoresis of AMP-deaminase isolated from the cirrhotic liver was performed. The obtained results indicate clearly that advanced proteolytic processes occurring in the cirrhotic liver may affect structural integrity of AMP-deaminase studied, making enzyme less active and less sensitive to regulatory action of important allosteric effectors.
Dutka
Paweł
P
Department of Anesthesiology and Intensive Therapy, Debinki Medical University of Gdansk, Gdansk, Poland.
Szydłowska
Magdalena
M
Chodorowski
Zygmunt
Z
Rybakowska
Iwona
I
Nagel-Starczynowska
Gabriela
G
Kaletha
Krystian
K
D000659:AMP Deaminase / Q000737:chemistry* / Q000378:metabolism*; D000328:Adult; D000494:Allosteric Regulation; D016022:Case-Control Studies; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008104:Liver Cirrhosis, Alcoholic / Q000201:enzymology*; D008297:Male; D008875:Middle Aged; D008970:Molecular Weight; D013379:Substrate Specificity
D000659:AMP Deaminase