An internal water-retention site in the rhomboid intramembrane protease GlpG ensures catalytic efficiency.

ABSTRACT: Rhomboid proteases regulate key cellular pathways, but their biochemical mechanism including how water is made available to the membrane-immersed active site remains ambiguous. We performed four prolonged molecular dynamics simulations initiated from both gate-open and gate-closed states of Escherichia coli rhomboid GlpG in a phospholipid bilayer. GlpG was notably stable in both gating states, experiencing similar tilt and local membrane thinning, with no observable gating transitions, highlighting that gating is rate-limiting. Analysis of dynamics revealed rapid loss of crystallographic waters from the active site, but retention of a water cluster within a site formed by His141, Ser181, Ser185, and/or Gln189. Experimental interrogation of 14 engineered mutants revealed an essential role f

SUBMITTER: Zhou Y 

PROVIDER: S-EPMC3571093 | biostudies-literature | 2012 Jul

REPOSITORIES: biostudies-literature