Generation and characterization of functional phosphoserine-incorporated neuronal nitric oxide synthase holoenzyme.

ABSTRACT: Phosphorylation is an important pathway for the regulation of nitric oxide synthase (NOS) at the posttranslational level. However, the molecular underpinnings of NOS regulation by phosphorylations remain unclear to date, mainly because of the problems in making a good amount of active phospho-NOS proteins. Herein, we have established a system in which recombinant rat nNOS holoprotein can be produced with site-specific incorporation of phosphoserine (pSer) at residue 1412, using a specialized bacterial host strain for pSer incorporation. The pSer1412 nNOS protein demonstrates UV-Vis, far-UV CD and fluorescence spectral properties that are identical to those of nNOS overexpressed in other bacterial strains. The protein is also functional, possessing normal NO production and NADPH oxidation a

SUBMITTER: Zheng H 

PROVIDER: S-EPMC6384131 | biostudies-literature | 2019 Feb

REPOSITORIES: biostudies-literature