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Mutational Analysis of the Dimer and Decamer Interface Residues in the Yeast Peroxiredoxin Tsa1
Author(s) -
Loberg Matthew,
Buchan John,
Allan Kristin,
Spencer Matthew,
West James
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.570.7
Subject(s) - peroxiredoxin , dimer , protein quaternary structure , cysteine , chemistry , yeast , biochemistry , mutant , peroxidase , gene , enzyme , organic chemistry , protein subunit
Oxidative damage of biological macromolecules is associated with a number of disease pathologies and the aging process. To defend against oxidative injury caused by peroxides, organisms express numerous peroxidases, including a family of conserved peroxiredoxins. Of the five peroxiredoxins found in baker's yeast, four of these proteins (Tsa1, Tsa2, Ahp1, and Prx1) are targets of the protein cross‐linker divinyl sulfone (DVSF) in treated yeast cells. While previous work has implicated the redox‐active cysteine residues in Tsa1 as targets of DVSF, the cysteine residues are brought into close proximity via dimeric quaternary structure. Therefore, we are investigating the quaternary structures formed by Tsa1, seeking to understand what amino acid residues are critical for the formation of dimers and higher order structures. To this end, we have performed a structure‐guided mutational analysis of conserved residues at the interfaces in Tsa1. Specifically, we have focused our studies on two putative ion‐pairing regions at the dimer interface and potential aromatic interactions at the decamer interface. Mutations at these sites in Tsa1 disrupt the quaternary structures formed, as measured by protein cross‐linking and native PAGE analysis. Moreover, the mutations studied compromise Tsa1′s ability to defend against oxidative insults and carry out other functions in cells.