Identification of the Binding Domain for NADP+ of Human Glucose-6-phosphate Dehydrogenase by Sequence Analysis of Mutants

Human erythrocyte glucose-6-phosphate dehydrogenase is normally quite stable in the presence of 10 μ M NADP+. Certain glucose-6-phosphate dehydrogenase variants lose virtually all their activity at this concentration of NADP+ but are reactivated by 200 μ M NADP+. Such variants presumably have a defe...

Full description

Saved in:
Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1989-12, Vol.86 (24), p.10015-10017
Main Authors: Hirono, Akira, Kuhl, Wanda, Gelbart, Terri, Forman, Linda, Fairbanks, Virgil F., Beutler, Ernest
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Amino acids
Analytical, structural and metabolic biochemistry
Animals
Binding Sites
Biochemistry
Biological and medical sciences
Complementary DNA
Dehydrogenases
DNA
DNA - genetics
Enzymes
Enzymes and enzyme inhibitors
Erythrocytes - enzymology
Exons
Fundamental and applied biological sciences. Psychology
Gene Amplification
Genes
Genetic mutation
Genetic Variation
Glucosephosphate Dehydrogenase - genetics
Humans
Molecular Sequence Data
Mutation
NADP - metabolism
Nucleotides
Oligonucleotide Probes
Oxidoreductases
Polymerase chain reaction
Sequence Homology, Nucleic Acid
Citations: Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Human erythrocyte glucose-6-phosphate dehydrogenase is normally quite stable in the presence of 10 μ M NADP+. Certain glucose-6-phosphate dehydrogenase variants lose virtually all their activity at this concentration of NADP+ but are reactivated by 200 μ M NADP+. Such variants presumably have a defect in their NADP+-binding site. We analyzed the sequence of cDNA or genomic DNA from seven unrelated patients with hemolytic anemia due to the inheritance of variants that are reactivated by NADP+. Six patients had substitutions of one of three adjacent amino acids, and the seventh patient had another amino acid substitution 23 residues downstream. These amino acids are highly conserved, all being present in rat and all but one being found also in Drosophila. The anomalous electrophoretic behavior of some of the variants can be explained by their loss of ability to bind NADP+. We conclude that the region in which these mutations occur defines the binding domain for NADP+ and that binding NADP+ that has been designated as ``structural'' and as ``catalytic'' probably occurs at the same site.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.86.24.10015