PMID- 29170897
OWN - NLM
STAT- MEDLINE
DCOM- 20190429
LR  - 20190429
IS  - 1432-1327 (Electronic)
IS  - 0949-8257 (Linking)
VI  - 23
IP  - 2
DP  - 2018 Mar
TI  - Selective tuning of activity in a multifunctional enzyme as revealed in the F21W 
      mutant of dehaloperoxidase B from Amphitrite ornata.
PG  - 209-219
LID - 10.1007/s00775-017-1520-x [doi]
AB  - Possessing both peroxidase and peroxygenase activities with a broad substrate 
      profile that includes phenols, indoles, and pyrroles, the enzyme dehaloperoxidase 
      (DHP) from Amphitrite ornata is a multifunctional catalytic hemoglobin that 
      challenges many of the assumptions behind the well-established structure-function 
      paradigm in hemoproteins. While previous studies have demonstrated that the F21W 
      variant leads to attenuated peroxidase activity in DHP, here we have studied the 
      impact of this mutation on peroxygenase activity to determine if it is possible 
      to selectively tune DHP to favor one function over another. Biochemical assays 
      with DHP B (F21W) revealed minimal decreases in peroxygenase activity of 
      1.2-2.1-fold as measured by 4-nitrophenol or 5-Br-indole substrate conversion, 
      whereas the peroxidase activity catalytic efficiency for 2,4,6-trichlorophenol 
      (TCP) was more than sevenfold decreased. Binding studies showed a 20-fold weaker 
      affinity for 5-bromoindole (K (d) = 2960 +/- 940 muM) in DHP B (F21W) compared to WT 
      DHP B. Stopped-flow UV/visible studies and isotope labeling experiments together 
      suggest that the F21W mutation neither significantly changes the nature of the 
      catalytic intermediates, nor alters the mechanisms that have been established for 
      peroxidase and peroxygenase activities in DHP. The X-ray crystal structure 
      (1.96 A; PDB 5VLX) of DHP B (F21W) revealed that the tryptophan blocks one of the 
      two identified TCP binding sites, specifically TCP(interior), suggesting that the 
      other site, TCP(exterior), remains viable for binding peroxygenase substrates. 
      Taken together, these studies demonstrate that blocking the TCP(interior) binding 
      site in DHP selectively favors peroxygenase activity at the expense of its 
      peroxidase activity.
FAU - Carey, Leiah M
AU  - Carey LM
AD  - Department of Chemistry, North Carolina State University, Raleigh, NC, 
      27695-8204, USA.
FAU - Kim, Kyung Beom
AU  - Kim KB
AD  - Department of Chemistry, North Carolina State University, Raleigh, NC, 
      27695-8204, USA.
AD  - Department of Fine Chemistry, Seoul National University of Science and 
      Technology, Seoul, 139-743, Korea.
FAU - McCombs, Nikolette L
AU  - McCombs NL
AD  - Department of Chemistry, North Carolina State University, Raleigh, NC, 
      27695-8204, USA.
FAU - Swartz, Paul
AU  - Swartz P
AD  - Department of Structural and Molecular Biochemistry, North Carolina State 
      University, Raleigh, NC, 27695-7622, USA.
FAU - Kim, Cheal
AU  - Kim C
AD  - Department of Fine Chemistry, Seoul National University of Science and 
      Technology, Seoul, 139-743, Korea.
FAU - Ghiladi, Reza A
AU  - Ghiladi RA
AUID- ORCID: 0000-0002-6450-9311
AD  - Department of Chemistry, North Carolina State University, Raleigh, NC, 
      27695-8204, USA. Reza_Ghiladi@ncsu.edu.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20171123
PL  - Germany
TA  - J Biol Inorg Chem
JT  - Journal of biological inorganic chemistry : JBIC : a publication of the Society 
      of Biological Inorganic Chemistry
JID - 9616326
RN  - 0 (Hemoglobins)
RN  - EC 1.11.1.- (DHP I dehaloperoxidase)
RN  - EC 1.11.1.- (Peroxidases)
SB  - IM
MH  - Animals
MH  - Catalysis
MH  - Crystallography, X-Ray
MH  - Hemoglobins/chemistry/genetics/isolation & purification/*metabolism
MH  - *Mutation
MH  - Peroxidases/chemistry/genetics/isolation & purification/*metabolism
MH  - Polychaeta/*enzymology
MH  - Spectrophotometry, Ultraviolet
MH  - Substrate Specificity
OTO - NOTNLM
OT  - Globin
OT  - Peroxidase
OT  - Peroxygenase
OT  - Structure-function relationship
EDAT- 2017/11/25 06:00
MHDA- 2019/04/30 06:00
CRDT- 2017/11/25 06:00
PHST- 2017/09/28 00:00 [received]
PHST- 2017/11/17 00:00 [accepted]
PHST- 2017/11/25 06:00 [pubmed]
PHST- 2019/04/30 06:00 [medline]
PHST- 2017/11/25 06:00 [entrez]
AID - 10.1007/s00775-017-1520-x [pii]
AID - 10.1007/s00775-017-1520-x [doi]
PST - ppublish
SO  - J Biol Inorg Chem. 2018 Mar;23(2):209-219. doi: 10.1007/s00775-017-1520-x. Epub 
      2017 Nov 23.