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Related Experiment Videos

A mutant T4 lysozyme displays five different crystal conformations.

H R Faber1, B W Matthews

  • 1Department of Physics, University of Oregon, Eugene 97403.

Nature
|November 15, 1990
PubMed
Summary

A T4 lysozyme mutant with isoleucine replacing methionine (Met 6) exhibits enhanced hinge-bending motion. This conformational flexibility was observed across five distinct crystal structures, without impacting enzyme activity or stability.

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Area of Science:

  • Biochemistry
  • Structural Biology
  • Molecular Biophysics

Background:

  • Phage T4 lysozyme possesses a two-domain structure with an active-site cleft.
  • Crystallographic data suggested hinge-bending motion in the wild-type enzyme's N-terminal domain.
  • This flexibility may facilitate substrate access to the active site.

Purpose of the Study:

  • To investigate the conformational flexibility of T4 lysozyme.
  • To characterize a mutant T4 lysozyme with enhanced hinge-bending motion.
  • To correlate structural changes with enzyme activity and stability.

Main Methods:

  • X-ray crystallography was employed to determine the structures of wild-type and mutant T4 lysozyme.
  • A mutant form (Met6Ile) was engineered and crystallized.

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  • Analysis of thermal displacement parameters and distinct molecular conformations in the crystal lattice.

    • Main Results:

    • The Met6Ile mutant T4 lysozyme crystallized in five different conformations.
    • Four independent molecules in one crystal form displayed significantly different structures.
    • The substitution enhanced the presumed hinge-bending motion of the enzyme.
    • Catalytic activity and thermal stability remained largely unaffected.

    Conclusions:

    • The Met6Ile substitution in T4 lysozyme amplifies hinge-bending motion.
    • Conformational flexibility is a key feature of T4 lysozyme.
    • Enhanced flexibility does not necessarily compromise enzymatic function or stability.