Mapping of a substrate binding site in the protein disulfide isomerase-related chaperone wind based on protein function and crystal structure.

The Journal of Biological Chemistry
Kathrin BarnewitzD M Ferrari

Abstract

The protein disulfide isomerase (PDI)-related protein Wind is essential in Drosophila melanogaster, and is required for correct targeting of Pipe, an essential Golgi transmembrane 2-O-sulfotransferase. Apart from a thioredoxin fold domain present in all PDI proteins, Wind also has a unique C-terminal D-domain found only in PDI-D proteins. Here, we show that Pipe processing requires dimeric Wind, which interacts directly with the soluble domain of Pipe in vitro, and we map an essential substrate binding site in Wind to the vicinity of an exposed cluster of tyrosines within the thioredoxin fold domain. In vitro, binding occurs to multiple sites within the Pipe polypeptide and shows specificity for two consecutive aromatic residues. A second site in Wind, formed by a cluster of residues within the D-domain, is likewise required for substrate processing. This domain, expressed separately, impairs Pipe processing by the full-length Wind protein, indicating competitive binding to substrate. Our data represent the most accurate map of a peptide binding site in a PDI-related protein available to date and directly show peptide specificity for a naturally occurring substrate.

References

Jun 1, 1983·European Journal of Biochemistry·O EppA Wendel
Aug 1, 1994·Trends in Biochemical Sciences·R B FreedmanM F Tuite
Jun 26, 1999·Journal of Biomolecular NMR·J KemminkN J Darby
May 8, 2002·The Journal of Biological Chemistry·Kevin G HoffLarry E Vickery
Oct 9, 2002·Genes & Development·Kazuyuki HoshijimaDavid Jonah Grunwald
Aug 23, 2003·The Journal of Biological Chemistry·Zhen ZhaoChih-Chen Wang
Aug 28, 2003·The Journal of Biological Chemistry·Qingjun MaDavid M Ferrari

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Citations

May 9, 2006·Antioxidants & Redox Signaling·Souren Mkrtchian, Tatyana Sandalova
May 30, 2009·Antioxidants & Redox Signaling·Feras Hatahet, Lloyd W Ruddock
Feb 3, 2007·Molecular Biology of the Cell·Emily K Rainey-BargerBilly Tsai
Nov 21, 2008·Journal of Virology·Emily K Rainey-BargerBilly Tsai
Dec 29, 2013·Apoptosis : an International Journal on Programmed Cell Death·Irina HirschDavid Michael Ferrari
Feb 5, 2013·Journal of Molecular Biology·Andreas FunknerDavid M Ferrari
Sep 26, 2007·The FEBS Journal·Feras Hatahet, Lloyd W Ruddock
Dec 29, 2005·Biochemical and Biophysical Research Communications·Mikhail BaryshevSouren Mkrtchian
Aug 30, 2014·Cell·Mattia R BordoliMalcolm Whitman
Dec 21, 2007·Biochimica Et Biophysica Acta·Christian Appenzeller-Herzog, Lars Ellgaard
Dec 17, 2008·Journal of Molecular Biology·Naomi N BarakDavid M Ferrari
Dec 14, 2004·The Journal of Biological Chemistry·Peppi KoivunenLloyd W Ruddock
Dec 2, 2004·The Journal of Biological Chemistry·Veronique M HermannMichael J Hubbard
Mar 17, 2019·The Journal of Biological Chemistry·Rajaiah PerguSivaram V S Mylavarapu
Apr 15, 2020·The FEBS Journal·Guennadi Kozlov, Kalle Gehring
Oct 6, 2020·Frontiers in Physiology·Margaret BreckerRonald C Rubenstein

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