Measured and calculated effects of mutations in bacteriophage T4 lysozyme on interactions in solution.
Understanding the molecular determinants of protein interactions in solution has fundamental implications for understanding protein solution thermodynamics and, hence, processes as diverse as separations performance and cellular self-organization. Our earlier theoretical calculations indicate that the protein-protein interactions are dominated by a small number of configurations in which highly complementary surface regions are apposed, rather than by the overall colloidal interactions. To examine this paradigm more explicitly, we investigated the effects of protein structural modifications on protein-protein interactions. Experimental measurements are presented of B(22)(') values of a set of mutants of Ser44 in bacteriophage T4 lysozyme. Effects are seen with both charged and uncharged substitutions. The results with the charged substitutions follow the expected trends, whereas those with the uncharged substitutions may be explained by the impact of the mutations on the local protein geometry, which directly affects the complementarity of protein interactions. These effects are also captured well by molecular calculations that account for the mutations. The interaction energetics between protein pairs could provide information on the propensity for adventitious interactions, which can have important implications for separations and for normal and pathological self-assembly. Thus, protein structural data implicit in genomic information, coupled with appropriate calculational and experimental tools, can ultimately provide insights into protein interactions in vivo and in bioprocessing.
Chang
R C
RC
Center for Molecular and Engineering Thermodynamics, Department of Chemical Engineering, University of Delaware, Newark, Delaware 19716, USA.
Asthagiri
D
D
Lenhoff
A M
AM
D017122:Bacteriophage T4 / Q000201:enzymology*; D008958:Models, Molecular; D009113:Muramidase / Q000737:chemistry* / Q000235:genetics; D009154:Mutation; D012996:Solutions
D012996:Solutions; D009113:Muramidase