2PQY

E. coli RNase 1 (in vitro refolded with DsbA only)

Summary for 2PQY

• Entry DOI: 10.2210/pdb2pqy/pdb
• Related: 2PQX
• Descriptor: Ribonuclease I, CALCIUM ION, 2-(N-MORPHOLINO)-ETHANESULFONIC ACID, ... (4 entities in total)
• Functional Keywords: ribonuclease, hydrolase
• Biological source: Escherichia coli
• Cellular location: Periplasm: P21338
• Total number of polymer chains: 1
• Total formula weight: 27428.01

Authors

Messens, J.,Loris, R. (deposition date: 2007-05-03, release date: 2007-08-14, Last modification date: 2024-10-30)

Primary citation

Messens, J.,Collet, J.F.,Van Belle, K.,Brosens, E.,Loris, R.,Wyns, L.
The oxidase DsbA folds a protein with a nonconsecutive disulfide.
J.Biol.Chem., 282:31302-31307, 2007

PubMed Abstract: One of the last unsolved problems of molecular biology is how the sequential amino acid information leads to a functional protein. Correct disulfide formation within a protein is hereby essential. We present periplasmic ribonuclease I (RNase I) from Escherichia coli as a new endogenous substrate for the study of oxidative protein folding. One of its four disulfides is between nonconsecutive cysteines. In general view, the folding of proteins with nonconsecutive disulfides requires the protein disulfide isomerase DsbC. In contrast, our study with RNase I shows that DsbA is a sufficient catalyst for correct disulfide formation in vivo and in vitro. DsbA is therefore more specific than generally assumed. Further, we show that the redox potential of the periplasm depends on the presence of glutathione and the Dsb proteins to maintain it at-165 mV. We determined the influence of this redox potential on the folding of RNase I. Under the more oxidizing conditions of dsb(-) strains, DsbC becomes necessary to correct non-native disulfides, but it cannot substitute for DsbA. Altogether, DsbA folds a protein with a nonconsecutive disulfide as long as no incorrect disulfides are formed.

PubMed: 17702751
DOI: 10.1074/jbc.M705236200

Experimental method

X-RAY DIFFRACTION (2.3 Å)