Triple point mutation Asp10-->His, Asn101-->Asp, Arg148-->Ser in T4 phage lysozyme leads to the molten globule

Protein Eng. 1992 Dec;5(8):781-3. doi: 10.1093/protein/5.8.781.

Abstract

The triple amino acid replacement (Asp10-->His, Asn101-->Asp, Arg148-->Ser) in T4 phage lysozyme was carried out by site-directed mutagenesis. At acid pH (2.7) the mutant is in a conformational state with the properties of the molten globule: (i) the mutant protein molecule is essentially compact; (ii) its CD spectrum in the near UV region is drastically reduced in intensity as compared with the wild type protein spectrum; (iii) the CD spectrum in the far UV region indicates the presence of pronounced secondary structure in the mutant; (iv) unlike the wild type protein the mutant protein can bind the hydrophobic fluorescent probe, ANS.

MeSH terms

  • Bacteriophage T4 / enzymology*
  • Bacteriophage T4 / genetics
  • Circular Dichroism
  • Fluorescent Dyes
  • Muramidase / chemistry*
  • Muramidase / genetics
  • Mutagenesis, Site-Directed
  • Protein Conformation
  • Protein Folding
  • Protein Structure, Secondary
  • Spectrophotometry, Ultraviolet

Substances

  • Fluorescent Dyes
  • Muramidase