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Probing solvation environments in Tt H‐NOX by site‐specific incorporation of 4‐cyano‐L‐phenylalanine
Author(s) -
Kearney Caroline,
Olenginski Lukasz T.,
Tariq Daniyal,
Hirn Trexler D.,
Brewer Scott H.,
PhillipsPiro Christine M.
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.603.15
Subject(s) - chemistry , heme , nox , nitric oxide , carbon monoxide , hemeprotein , phenylalanine , amino acid , biochemistry , stereochemistry , catalysis , organic chemistry , enzyme , combustion
Heme Nitric Oxide and/or Oxygen (H‐NOX) binding proteins are a diverse group of hemoproteins that bind diatomic gases like nitric oxide (NO), carbon monoxide (CO), and/or oxygen (O 2 ). H‐NOX proteins, which can be found in both prokaryotic and eukaryotic cells, contain two distinct domains: a heme‐containing sensor domain that binds gaseous ligands and an effector domain that generates a cascade of signals for cells to function. Here local protein environments in Thermoanaerobacter tengcongensis H‐NOX ( Tt H‐NOX) were investigated using the vibrational reporter unnatural amino acid (UAA) 4‐cyano‐L‐phenylalanine (pCNF). This UAA was genetically site‐specifically incorporated into Tt H‐NOX at multiple positions individually including near the heme pocket using amber codon suppression technology. Infrared analysis of the constructs probing the position of the nitrile symmetric stretch of pCNF revealed a myriad of local environments in the protein. These results highlighting solvated or buried positions in the protein will be presented. Support or Funding Information NIH R15GM1219842CMPP and the Fred A. Snavely Research Award