Affinity labeling of a lysine residue in the coenzyme binding site of pig heart mitochondrial malate dehydrogenase

Biochemistry ◽  
1979 ◽  
Vol 18 (21) ◽  
pp. 4683-4690 ◽  
Author(s):  
Siddhartha Roy ◽  
Roberta F. Colman
Keyword(s):  
Binding Site ◽  
Malate Dehydrogenase ◽  
Lysine Residue ◽  
Affinity Labeling ◽  
Coenzyme Binding ◽  
Mitochondrial Malate Dehydrogenase ◽  
Coenzyme Binding Site

Notizen: Coenzyme Binding at Different Ionization States of Cytoplasmic and Mitochondrial Malate Dehydrogenase

1982 ◽  
Vol 37 (5-6) ◽  
pp. 547-549 ◽  
Author(s):  
Klaus Schwerdtfeger ◽  
Christoph Woenckhaus ◽  
David M. Parker ◽  
John J. Holbrook
Keyword(s):  
Binding Site ◽  
Malate Dehydrogenase ◽  
Lower Limit ◽  
Mitochondrial Enzyme ◽  
Binding Studies ◽  
Active Centre ◽  
Cytoplasmic Enzyme ◽  
Coenzyme Binding ◽  
Mitochondrial Malate Dehydrogenase ◽  
Ionizable Groups

Abstract pH-titrations with NADH show two ionizable groups in mitochondrial and cytoplasmic malate dehydrogenase, the first with a pKa in the range 6.8 -8.3 for the mitochondrial and 6.4-7.8 for the cytoplasmic enzyme, the second with a lower limit at 10.2 resp. 11. Comparison with bis-(dihydronicotinamide)-dinucleotide and dihydronicotina-mide-ribosyl-P2-ribose-pyrophosphate instead of NADH indicates that the second alkaline ionization is caused by a residue placed near the adenine binding site of the active centre of the two isoenzymes. Binding studies with NADH and NAD+ give evidence for the participation of a group in the mitochondrial enzyme with pKa 6.8, deprotonation of which is necessary for detectable association of NAD+. In contrast the fixation of NAD+ to the cytoplasmic enzyme is independent of pH.

scholarly journals Affinity labelling of the NADP+-binding site of glucose 6-phosphate dehydrogenase from Candida utilis

1979 ◽  
Vol 183 (2) ◽  
pp. 297-302 ◽  
Author(s):  
T Bellini ◽  
M Signorini ◽  
F Dallocchio ◽  
M Rippa
Keyword(s):  
Binding Site ◽  
Candida Utilis ◽  
Aldehyde Group ◽  
Lysine Residue ◽  
Reversible Inactivation ◽  
Coenzyme Binding ◽  
Complete Inactivation ◽  
Affinity Labelling ◽  
Glucose 6 Phosphate Dehydrogenase ◽  
Coenzyme Binding Site

1. Periodate-oxidized NADP+ inhibits the catalytic activity of glucose 6-phosphate dehydrogenase from Candida utilis, competing with NADP+. 2. Incubation of the enzyme with the coenzyme analogue causes partial reversible inactivation of the enzyme as a result of affinity labelling of the coenzyme-binding site. 3. Some kinetic values of the reaction were calculated. 4. The inactivation can be made irreversible by treatment with NaBH4, which reduces a Schiff base formed between an aldehyde group on the coenzyme analogue and a lysine residue on the enzyme. 5. Complete inactivation can be correlated with the binding of only one inhibitor to each enzyme subunit. 6. The lysine residue involved in the binding of the inhibitor is present at the coenzyme-binding site.

scholarly journals Study of Coenzyme Binding Site of Octopine Dehydrogenase Using Analogues of NAD+

1975 ◽  
Vol 56 (1) ◽  
pp. 109-116 ◽  
Author(s):  
Anna OLOMUCKI ◽  
Francoise THOME-BEAU ◽  
Jean-Francois BIELLMANN ◽  
Guy BRANLANT
Keyword(s):  
Binding Site ◽  
Coenzyme Binding ◽  
Octopine Dehydrogenase ◽  
Coenzyme Binding Site

scholarly journals Evidence for an arginine residue at the coenzyme-binding site of Escherichia coli isocitrate dehydrogenase

1989 ◽  
Vol 261 (1) ◽  
pp. 301-304 ◽  
Author(s):  
J S McKee ◽  
H G Nimmo
Keyword(s):  
Escherichia Coli ◽  
Binding Site ◽  
Isocitrate Dehydrogenase ◽  
Arginine Residue ◽  
Order Process ◽  
First Order ◽  
Coenzyme Binding ◽  
Phosphorylated Form ◽  
Specific Reagent ◽  
Coenzyme Binding Site

The arginine-specific reagent phenylglyoxal inactivated the active, dephosphorylated, form of Escherichia coli isocitrate dehydrogenase rapidly in a pseudo-first-order process. Both NADP+ and NADPH protected the enzyme against inactivation. Phenylglyoxal appeared to react with one arginine residue per subunit, and the extent of the reaction was proportional to the extent of the inactivation. In contrast, the phosphorylated form of isocitrate dehydrogenase did not react detectably with phenylglyoxal. The data indicate that the coenzyme-binding site of isocitrate dehydrogenase contains a reactive arginine residue that is protected by phosphorylation, and are consistent with the hypothesis that phosphorylation of the enzyme occurs close to or at its active site.

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