Functional and structural role of a tryptophan generally observed in protein-carbohydrate interaction. TRP-62 of hen egg white lysozyme

J Biol Chem. 1994 Mar 11;269(10):7070-5.

Abstract

In order to elucidate the role of the aromatic ring in recognition of the sugar ring, Trp-62 of hen egg white lysozyme, which is proposed on the basis of x-ray crystallography data to make contact with a sugar ring through van der Waals interaction, was replaced with aliphatic amino acids (Leu, Ile, Val, and Ala) and Gly by site-directed mutagenesis. In spite of the loss of the aromatic effect, these mutant lysozymes, except for the Trp-62-->Gly mutant, showed higher bacteriolytic activity than the wild-type lysozyme. Furthermore, the Trp-62-->Gly mutant still retained appreciable bacteriolytic activity. On the other hand, by these replacements, the enzymatic activities toward non-charged substrates were markedly reduced. Additionally, the side-chain structure of position 62 was found to be largely responsible for recognition of a saccharide ring in its active site cleft. NMR analysis of the Trp-62-->Leu and Trp-62-->Gly mutants indicated that the structural effects of Trp-62 replacements were localized in the loop region around position 62 and the part of the beta-sheet containing the hydrogen bonding network important for enzymatic activity. Thus, we conclude that Trp-62 not only interacts with oligosaccharide through van der Waals contact, but also maintains the local structural conformation to produce the lysozyme-oligosaccharide interaction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Binding Sites
  • Carbohydrate Metabolism*
  • Carbohydrate Sequence
  • Chickens
  • Glycine / chemistry
  • Magnetic Resonance Spectroscopy
  • Muramidase / chemistry
  • Muramidase / genetics
  • Muramidase / metabolism*
  • Mutation
  • Protein Conformation
  • Tryptophan / chemistry
  • Tryptophan / physiology*

Substances

  • Tryptophan
  • Muramidase
  • Glycine