An enzymatic method for precise oxygen affinity measurements over nanomolar-to-millimolar concentration regime.

ABSTRACT: Oxygen affinity is an important property of metalloproteins that helps elucidate their reactivity profile and mechanism. Heretofore, oxygen affinity values were determined either using flash photolysis and polarography techniques that require expensive instrumentation, or using oxygen titration methods which are erroneous at low nanomolar and at high millimolar oxygen concentrations. Here, we describe an inexpensive, easy-to-setup, and a one-pot method for oxygen affinity measurements that uses the enzyme chlorite dismutase (Cld) as a precise in situ oxygen source. Using this method, we measure thermodynamic and kinetic oxygen affinities (K_d and K_M) of different classes of heme and non-heme metalloproteins involved in oxygen transport, sensing, and catalysis. The meth

SUBMITTER: Sanyal R 

PROVIDER: S-EPMC9153948 | biostudies-literature | 2020 Mar

REPOSITORIES: biostudies-literature