BIOMAT Database Logo BIOMAT Database Logo

Interaction of Leuconostoc mesenteroides glucose-6-phosphate dehydrogenase with pyridoxal 5'-diphospho-5'-adenosine. Affinity labeling of Lys-21 and Lys-343. 1991

J R LaDine, and D Carlow, and W T Lee, and R L Cross, and T G Flynn, and H R Levy
Department of Biochemistry and Molecular Biology, State University of New York Health Science Center, Syracuse 13210.

Pyridoxal 5'-diphospho-5'-adenosine (PLP-AMP) inhibits glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides competitively with respect to glucose 6-phosphate and noncompetitively with respect to NAD+ or NADP+, with Ki = 40 microM in the NADP-linked and 34 microM in the NAD-linked reaction. Incubation of glucose-6-phosphate dehydrogenase with [3H]PLP-AMP followed by borohydride reduction shows that incorporation of 0.85 mol of PLP-AMP per mol of enzyme subunit is required for complete inactivation. Both glucose 6-phosphate and NAD+ protect against this covalent modification. The proteolysis of the modified enzyme and isolation and sequencing of the labeled peptides revealed that Lys-21 and Lys-343 are the sites of PLP-AMP interaction and that glucose 6-phosphate and NAD+ protect both lysyl residues against modification. Pyridoxal 5'-phosphate (PLP) also modifies Lys-21 and probably Lys-343. Lys-21 is part of a highly conserved region that is present in all glucose-6-phosphate dehydrogenases that have been sequenced. Lys-343 corresponds to an arginyl residue in other glucose-6-phosphate dehydrogenases and is in a region that is less homologous with those enzymes. PLP-AMP and PLP are believed to interact with L. mesenteroides glucose-6-phosphate dehydrogenase at the glucose 6-phosphate binding site. Simultaneous binding of NAD+ induces conformational changes (Kurlandsky, S. B., Hilburger, A. C., and Levy, H. R. (1988) Arch. Biochem. Biophys. 264, 93-102) that are postulated to interfere with Schiff's-base formation with PLP or PLP-AMP. One or both of the lysyl residues covalently modified by PLP or PLP-AMP may be located in regions of the enzyme undergoing the NAD(+)-induced conformational changes.

UI MeSH Term Description Entries
D007700 Kinetics The rate dynamics in chemical or physical systems.
D007934 Leuconostoc A genus of gram-positive, facultatively anaerobic bacteria whose growth is dependent on the presence of a fermentable carbohydrate. It is nonpathogenic to plants and animals, including humans. Leukonostoc
D011732 Pyridoxal Phosphate This is the active form of VITAMIN B 6 serving as a coenzyme for synthesis of amino acids, neurotransmitters (serotonin, norepinephrine), sphingolipids, aminolevulinic acid. During transamination of amino acids, pyridoxal phosphate is transiently converted into pyridoxamine phosphate (PYRIDOXAMINE). Pyridoxal 5-Phosphate,Pyridoxal-P,Phosphate, Pyridoxal,Pyridoxal 5 Phosphate,Pyridoxal P
D002851 Chromatography, High Pressure Liquid Liquid chromatographic techniques which feature high inlet pressures, high sensitivity, and high speed. Chromatography, High Performance Liquid,Chromatography, High Speed Liquid,Chromatography, Liquid, High Pressure,HPLC,High Performance Liquid Chromatography,High-Performance Liquid Chromatography,UPLC,Ultra Performance Liquid Chromatography,Chromatography, High-Performance Liquid,High-Performance Liquid Chromatographies,Liquid Chromatography, High-Performance
D005954 Glucosephosphate Dehydrogenase Glucose-6-Phosphate Dehydrogenase,Dehydrogenase, Glucose-6-Phosphate,Dehydrogenase, Glucosephosphate,Glucose 6 Phosphate Dehydrogenase
D000244 Adenosine Diphosphate Adenosine 5'-(trihydrogen diphosphate). An adenine nucleotide containing two phosphate groups esterified to the sugar moiety at the 5'-position. ADP,Adenosine Pyrophosphate,Magnesium ADP,MgADP,Adenosine 5'-Pyrophosphate,5'-Pyrophosphate, Adenosine,ADP, Magnesium,Adenosine 5' Pyrophosphate,Diphosphate, Adenosine,Pyrophosphate, Adenosine
D000345 Affinity Labels Analogs of those substrates or compounds which bind naturally at the active sites of proteins, enzymes, antibodies, steroids, or physiological receptors. These analogs form a stable covalent bond at the binding site, thereby acting as inhibitors of the proteins or steroids. Affinity Labeling Reagents,Labeling Reagents, Affinity,Labels, Affinity,Reagents, Affinity Labeling
D013379 Substrate Specificity A characteristic feature of enzyme activity in relation to the kind of substrate on which the enzyme or catalytic molecule reacts. Specificities, Substrate,Specificity, Substrate,Substrate Specificities

Related Publications

J R LaDine, and D Carlow, and W T Lee, and R L Cross, and T G Flynn, and H R Levy
Glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides.
April 1989, Biochemical Society transactions,
J R LaDine, and D Carlow, and W T Lee, and R L Cross, and T G Flynn, and H R Levy
Glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides.
January 1975, Methods in enzymology,
J R LaDine, and D Carlow, and W T Lee, and R L Cross, and T G Flynn, and H R Levy
A glucose-6-phosphate dehydrogenase in Leuconostoc mesenteroides.
July 1953, Journal of bacteriology,
J R LaDine, and D Carlow, and W T Lee, and R L Cross, and T G Flynn, and H R Levy
Glucose 6-phosphate dehydrogenase from Leuconostoc mesenteroides. Kinetic studies.
April 1971, The Journal of biological chemistry,
J R LaDine, and D Carlow, and W T Lee, and R L Cross, and T G Flynn, and H R Levy
Lysine-21 of Leuconostoc mesenteroides glucose 6-phosphate dehydrogenase participates in substrate binding through charge-charge interaction.
March 1992, Protein science : a publication of the Protein Society,
J R LaDine, and D Carlow, and W T Lee, and R L Cross, and T G Flynn, and H R Levy
The adenine nucleotide binding site on yeast hexokinase PII. Affinity labeling of Lys-111 by pyridoxal 5'-diphospho-5'-adenosine.
June 1988, The Journal of biological chemistry,
J R LaDine, and D Carlow, and W T Lee, and R L Cross, and T G Flynn, and H R Levy
Glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides. Interaction of the enzyme with coenzymes and coenzyme analogs.
November 1976, Archives of biochemistry and biophysics,
J R LaDine, and D Carlow, and W T Lee, and R L Cross, and T G Flynn, and H R Levy
Aldose reductase from human psoas muscle. Affinity labeling of an active site lysine by pyridoxal 5'-phosphate and pyridoxal 5'-diphospho-5'-adenosine.
February 1989, The Journal of biological chemistry,
J R LaDine, and D Carlow, and W T Lee, and R L Cross, and T G Flynn, and H R Levy
Glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides: ligand-induced conformational changes.
July 1988, Archives of biochemistry and biophysics,
J R LaDine, and D Carlow, and W T Lee, and R L Cross, and T G Flynn, and H R Levy
The adenine nucleotide-binding site on yeast 3-phosphoglycerate kinase. Affinity labeling of Lys-131 by pyridoxal 5'-diphospho-5'-adenosine.
April 1991, The Journal of biological chemistry,
Copied contents to your clipboard!