Isolation and in Vitro Phosphorylation of Sensory Transduction Components Controlling Anaerobic Induction of Light Harvesting and Reaction Center Gene Expression in Rhodobacter capsulatus

Anaerobic induction of light harvesting and reaction center gene expression involves two transacting factors termed RegA and RegB. Sequence and mutational analysis has indicated that RegA and RegB constitute cognate components of a prokaryotic sensory transduction cascade with RegB comprising a memb...

Full description

Saved in:
Bibliographic Details
Published in:Biochemistry (Easton) 1995-01, Vol.34 (2), p.391-396
Main Authors: Inoue, Kazuhito, Kouadio, Jean-louis K, Mosley, Chere S, Bauer, Carl E
Format: Article
Language:English
Subjects:
Bacterial Proteins - genetics
Bacterial Proteins - isolation & purification
Bacterial Proteins - metabolism
Base Sequence
DNA Primers
Escherichia coli
Molecular Sequence Data
Phosphorylation
Photosynthetic Reaction Center Complex Proteins - genetics
Photosynthetic Reaction Center Complex Proteins - isolation & purification
Photosynthetic Reaction Center Complex Proteins - metabolism
Protein Folding
Protein Kinases
Recombinant Proteins
Rhodobacter capsulatus
Rhodobacter capsulatus - chemistry
Rhodobacter capsulatus - metabolism
Citations: Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Anaerobic induction of light harvesting and reaction center gene expression involves two transacting factors termed RegA and RegB. Sequence and mutational analysis has indicated that RegA and RegB constitute cognate components of a prokaryotic sensory transduction cascade with RegB comprising a membrane-spanning sensor kinase and RegA a cytosolic response regulator. In this study we have purified RegA, as well as a truncated portion of RegB (RegB') and undertaken an in vitro analysis of autophosphorylation and phosphotransfer activities. Incubation of RegB' with [gamma-32P]ATP and MgCl2 resulted in phosphorylation of RegB' (RegB' approximately P) over a 20-min incubation period. Incubation of RegB' approximately P with RegA resulted in rapid transfer of the phosphate from RegB' to RegA. In analogy to other characterized prokaryotic sensory transduction components, mutational and chemical stability studies also indicate that RegB' is autophosphorylated at a conserved histidine and that RegA accepts the phosphate from RegB at a conserved aspartate.
ISSN:0006-2960
1520-4995
DOI:10.1021/bi00002a002