Molecular Model of Human β-Cell Glucokinase Built by Analogy to the Crystal Structure of Yeast Hexokinase B

  1. Irene T Weber
  1. Department of Pharmacology, Jefferson Cancer Institute, Thomas Jefferson University Philadelphia, Pennsylvania
  2. Howard Hughes Medical Institute and Departments of Biochemistry and Molecular Biology and Medicine, University of Chicago Illinois
  3. Department of Physiology and Biophysics, State University of New York Stony Brook, New York
  1. Address correspondence and reprint requests to Dr. Irene T. Weber, Department of Pharmacology, Jefferson Cancer Institute, Thomas Jefferson University, 233 South 10th Street, Philadelphia, PA 19107.

Abstract

Recent studies have shown that mutations in human β-cell glucokinase that impair the activity of this key regulatory enzyme of glycolysis can cause early-onset non-insulin-dependent diabetes mellitus (NIDDM). The amino acid sequence of human glucokinase has 31% identity with yeast hexokinase, a related enzyme for which the crystal structure has been determined. This homology has allowed us to model the three-dimensional structure of human glucokinase by analogy to the crystal structure of yeast hexokinase B. This model of human glucokinase provides a basis for understanding the effects of mutations on its enzymatic activity. Residues in the active site and on the surface of the binding cleft for glucose are highly conserved in both enzymes. Regions far from the active site are predicted to differ in conformation, and 10 insertions or deletions that range in size from 1 to 7 residues are located on the protein surface between elements of secondary structure. The model structure suggests that human glucokinase binds glucose in a similar manner to yeast hexokinase. The glucose-binding site contains a conserved aspartic acid, two conserved glutamic acids, and two conserved asparagines that form hydrogen bond interactions with the hydroxyls of the glucose similar to those observed in other sugar-binding proteins. Mutation of residues in the predicted glucose-binding site has been found to greatly reduce enzymatic activity. This model will be useful for future structure/function studies of glucokinase.

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