The pK_a values of the catalytic residues in the retaining glycoside hydrolase T26H mutant of T4 lysozyme.

ABSTRACT: T4 phage lysozyme (T4L) is an enzyme that cleaves bacterial cell wall peptidoglycan. Remarkably, the single substitution of the active site Thr26 to a His (T26H) converts T4L from an inverting to a retaining glycoside hydrolase with transglycosylase activity. It has been proposed that T26H-T4L follows a double displacement mechanism with His26 serving as a nucleophile to form a covalent glycosyl-enzyme intermediate (Kuroki et al., PNAS 1999; 96:8949-8954). To gain further insights into this or alternative mechanisms, we used NMR spectroscopy to measure the acid dissociation constants (pK_a values) and/or define the ionization states of the Asp, Glu, His, and Arg residues in the T4L mutant. Most notably, the pK_a value of the putative nucleophile His26 is 6.8 ± 0.1, wher

SUBMITTER: Brockerman JA 

PROVIDER: S-EPMC6371219 | biostudies-literature | 2019 Mar

REPOSITORIES: biostudies-literature