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Science 23 January 2004:
Vol. 303. no. 5657, pp. 534 - 537
DOI: 10.1126/science.1091724
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Reports

Snapshots of DsbA in Action: Detection of Proteins in the Process of Oxidative Folding

Hiroshi Kadokura,1 Hongping Tian,1* Thomas Zander,2{dagger} James C. A. Bardwell,2 Jon Beckwith1{ddagger}

DsbA, a thioredoxin superfamily member, introduces disulfide bonds into newly translocated proteins. This process is thought to occur via formation of mixed disulfide complexes between DsbA and its substrates. However, these complexes are difficult to detect, probably because of their short-lived nature. Here we show that it is possible to detect such covalent intermediates in vivo by a mutation in DsbA that alters cis proline-151. Further, this mutant allowed us to identify substrates of DsbA. Alteration of the cis proline, highly conserved among thioredoxin superfamily members, may be useful for the detection of substrates and intermediate complexes in other systems.

1 Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, MA 02115, USA.
2 Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, MI 48109, USA.



* Present address: Harvard School of Public Health, Boston, MA 02115, USA.

{dagger} Present address: Profos Ag, Josef-Engert-Straße 9, 93053 Regensburg, Germany.

{ddagger} To whom correspondence should be addressed. E-mail: jbeckwith{at}hms.harvard.edu

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