Download An additional cysteine in a typical 2

An additional cysteine in a typical 2-Cys peroxiredoxin of
Pseudomonas promotes functional switching between peroxidase and
molecular chaperone
Byung Chull Ana, Seung Sik Leea, Hyun Suk Jungb, Jin Young Kimc, Yuno Leed, Keun Woo
Leed, Sang Yeol Leed, Bhumi Nath Tripathia and Byung Yeoup Chunga,*
a
Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute, 29
Geumgu-gil, Jeongeup 580-185, Korea.
b
Department of Biochemistry, College of Natural Sciences, Kangwon National University,
Chuncheon 200-701, Korea.
c
Division of Mass Spectrometry Research, Korea Basic Science Institute, 162 Yeongudanjiro,
Ochang 363-883, Korea.
d
Division of Applied Life Science (Brain Korea 21 Program), Gyeongsang National University,
501 Jinju-daero, Jinju 660-701, Korea.
Running title: Functional and structural change of peroxiredoxin.
* Corresponding author at: Advanced Radiation Technology Institute, Korea Atomic Energy
Research Institute, 29 Geumgu-gil, Jeongeup 580-185, Korea. Fax: +82 63 570 3331.
E-mail address: [email protected] (B.Y. Chung)
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Figure legends
Fig. S1. Amino acid sequence alignment and comparison of Prxs. Alignment of the amino acid
sequences of P. putida PpPrx (2-Cys Prx) with homologous AhpC and Prxs from several
representative prokaryotes and eukaryotes. The gaps (dashes) were introduced to optimize
sequence alignment. The highly conserved tripeptide Val-Cys-Pro (VCP), related to the catalytic
function, is designated with a square, and the additional cysteine (Cys112) is designated with a
triangle. ‘*’ indicates positions which have a single, fully conserved residue. ‘:’ indicates that
one of the following ‘strong’ groups is fully conserved. ‘.’ indicates that one of the following
‘weaker’ groups is fully conserved. The abbreviations for the amino acid sequences of Prxs
from various species are as follows: PpPrx, P. putida KT2440; PaPrx, P. aeruginosa PAO1;
PopPrx, Porphyra purpurea; LiPrx, Leptospira interrogans; PiPrx, Phytophthora infestans;
RnPrx, Rattus norvegicus; EncPrx, Encephalitozoon cuniculi GB-M1; EfAhpC, Enterococcus
faecalis V583; PbAhpC, Propionibacterium.
Fig. S2. Each SEC fraction was separated by 10% native PAGE and then visualized by silver
staining. F-1 fraction of PpPrx (lane 1), F-2 fraction of PpPrx (lane 2), F-1 fraction of PaPrx
(lane 3), and F-2 fraction of PaPrx (lane 4).
Fig. S3. MS and MS/MS spectra of tryptic digests with DTT and IAA treatment. (A)
YTLAADMTHEICcamK (Cys112, theoretical doubly charged monoisotopic molecular ion: m/z
776.85) from WT-PpPrx. (B) YTLAADTKHEICcamK (Cys112, theoretical doubly charged
monoisotopic molecular ion: m/z 775.38) from A112C-PaPrx. Manual assignment of MS/MS
spectra show fragments of peptide backbone and characteristic fragments of disulfide linkage,
including cysteine persulfide (monomer peptide +32 Da) and dehydroalanine (monomer peptide
-34 Da).
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Fig. S1
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Fig. S2
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Fig. S3
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