Protein substrate discrimination in the quiescin sulfhydryl oxidase (QSOX) family.

Biochemistry
Jennifer A CoddingColin Thorpe

Abstract

This work explores the substrate specificity of the quiescin sulfhydryl oxidase (QSOX) family of disulfide-generating flavoenzymes to provide enzymological context for investigation of the physiological roles of these facile catalysts of oxidative protein folding. QSOX enzymes are generally unable to form disulfide bonds within well-structured proteins. Use of a temperature-sensitive mutant of ubiquitin-conjugating enzyme 4 (Ubc4') as a model substrate shows that QSOX activity correlates with the unfolding of Ubc4' monitored by circular dichroism. Fusion of Ubc4' with the more stable glutathione-S-transferase domain demonstrates that QSOX can selectively introduce disulfides into the less stable domain of the fusion protein. In terms of intermolecular disulfide bond generation, QSOX is unable to cross-link well-folded globular proteins via their surface thiols. However, the construction of a septuple mutant of RNase A, retaining a single cysteine residue, demonstrates that flexible protein monomers can be directly coupled by the oxidase. Steady- and pre-steady-state kinetic experiments, combined with static fluorescence approaches, indicate that while QSOX is an efficient catalyst for disulfide bond formation between mobile ele...Continue Reading

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Citations

Feb 19, 2014·Biochimica Et Biophysica Acta·Kerstin Kojer, Jan Riemer
Apr 12, 2016·Brain Research·Sonam Parakh, Julie D Atkin
Jan 24, 2014·Biochemical Society Transactions·Rachel E MartinNeil J Bulleid
Jan 29, 2014·Free Radical Biology & Medicine·Benjamin A IsraelColin Thorpe
May 30, 2013·The Biochemical Journal·Jana RudolfNeil J Bulleid
Mar 30, 2016·Annual Review of Chemical and Biomolecular Engineering·Michael C Yi, Chaitan Khosla
Jan 13, 2015·Biological Chemistry·Marcel Deponte, Christopher Horst Lillig
Jul 13, 2017·Chemical Reviews·Deborah Fass, Colin Thorpe
May 3, 2018·Antioxidants & Redox Signaling·Daniela MennerichThomas Kietzmann
Apr 27, 2018·Protein Science : a Publication of the Protein Society·Tiantian YuColin Thorpe
Dec 14, 2016·Aging·Yi-An ZhanWen-Quan Zou
Jul 3, 2019·Frontiers in Cell and Developmental Biology·Sakari Kellokumpu

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