Abstract
An essential histidine ligand to the electron transfer copper (Cu H) of peptidylglycine α-hydroxylating monooxygenase (PHMcc) was mutated to an alanine and found to retain copper binding and hydroxylase activity [Jaron, S., et al. (2002) Biochemistry 41, 13274-13282]. An extensive kinetic and deuterium isotope effect study finds this mutant to maintain full coupling of O2 consumed to product formed despite a 3 order-of-magnitude decrease in kcat and a 300-fold decrease in k cat/Km(O2). Unexpectedly, electron transfer is not rate-limiting in H172A. Rather, the increased kinetic isotope effect (KIE) on kcat of 3.27 ± 0.39 suggests that C-H bond cleavage has become more rate-limiting, implicating a role for His172 that goes beyond that of a simple ligand to CUH. The mechanistic implications are discussed.
Original language | English (US) |
---|---|
Pages (from-to) | 15419-15429 |
Number of pages | 11 |
Journal | Biochemistry |
Volume | 45 |
Issue number | 51 |
DOIs | |
State | Published - Dec 26 2006 |
ASJC Scopus subject areas
- Biochemistry