Thioredoxin Profiling of Multiple Thioredoxin-Like Proteins in Staphylococcus aureus
Hydrogen sulfide (H2S) is thought to signal through protein S-sulfuration (persulfidation; S-sulfhydration) in both mammalian systems and bacteria. We previously profiled proteome S-sulfuration in Staphylococcus aureus (S. aureus) and identified two thioredoxin-like proteins, designated TrxP and Trx...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2018-10-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fmicb.2018.02385/full |
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| author | Hui Peng Hui Peng Yixiang Zhang Yixiang Zhang Jonathan C. Trinidad Jonathan C. Trinidad David P. Giedroc David P. Giedroc |
| author_facet | Hui Peng Hui Peng Yixiang Zhang Yixiang Zhang Jonathan C. Trinidad Jonathan C. Trinidad David P. Giedroc David P. Giedroc |
| author_sort | Hui Peng |
| collection | DOAJ |
| description | Hydrogen sulfide (H2S) is thought to signal through protein S-sulfuration (persulfidation; S-sulfhydration) in both mammalian systems and bacteria. We previously profiled proteome S-sulfuration in Staphylococcus aureus (S. aureus) and identified two thioredoxin-like proteins, designated TrxP and TrxQ, that were capable of reducing protein persulfides as a potential regulatory mechanism. In this study, we further characterize TrxP, TrxQ and the canonical thioredoxin, TrxA, by identifying candidate protein substrates in S. aureus cells using a mechanism-based profiling assay where we trap mixed disulfides that exist between the attacking cysteine of a FLAG-tagged Trx and a persulfidated cysteine on the candidate substrate protein in cells. Largely non-overlapping sets of four, 32 and three candidate cellular substrates were detected for TrxA, TrxP, and TrxQ, respectively, many of which were previously identified as global proteome S-sulfuration targets including for example, pyruvate kinase, PykA. Both TrxA (kcat = 0.13 s-1) and TrxP (kcat = 0.088 s-1) are capable of reducing protein persulfides on PykA, a model substrate detected as a candidate substrate of TrxP; in contrast, TrxQ shows lower activity (kcat = 0.015 s-1). This work reveals that protein S-sulfuration, central to H2S and reactive sulfur species (RSS) signaling, may impact cellular activities and appears to be regulated in S. aureus largely by TrxP under conditions of sulfide stress. |
| first_indexed | 2024-12-23T05:57:08Z |
| format | Article |
| id | doaj.art-83b25c1230464ee28475cc8fde7b0f4f |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-23T05:57:08Z |
| publishDate | 2018-10-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-83b25c1230464ee28475cc8fde7b0f4f2022-12-21T17:57:46ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2018-10-01910.3389/fmicb.2018.02385410891Thioredoxin Profiling of Multiple Thioredoxin-Like Proteins in Staphylococcus aureusHui Peng0Hui Peng1Yixiang Zhang2Yixiang Zhang3Jonathan C. Trinidad4Jonathan C. Trinidad5David P. Giedroc6David P. Giedroc7Department of Chemistry, Indiana University Bloomington, Bloomington, IN, United StatesBiochemistry Graduate Program, Indiana University Bloomington, Bloomington, IN, United StatesDepartment of Chemistry, Indiana University Bloomington, Bloomington, IN, United StatesLaboratory for Biological Mass Spectrometry, Department of Chemistry, Indiana University Bloomington, Bloomington, IN, United StatesDepartment of Chemistry, Indiana University Bloomington, Bloomington, IN, United StatesLaboratory for Biological Mass Spectrometry, Department of Chemistry, Indiana University Bloomington, Bloomington, IN, United StatesDepartment of Chemistry, Indiana University Bloomington, Bloomington, IN, United StatesDepartment of Molecular and Cellular Biochemistry, Indiana University Bloomington, Bloomington, IN, United StatesHydrogen sulfide (H2S) is thought to signal through protein S-sulfuration (persulfidation; S-sulfhydration) in both mammalian systems and bacteria. We previously profiled proteome S-sulfuration in Staphylococcus aureus (S. aureus) and identified two thioredoxin-like proteins, designated TrxP and TrxQ, that were capable of reducing protein persulfides as a potential regulatory mechanism. In this study, we further characterize TrxP, TrxQ and the canonical thioredoxin, TrxA, by identifying candidate protein substrates in S. aureus cells using a mechanism-based profiling assay where we trap mixed disulfides that exist between the attacking cysteine of a FLAG-tagged Trx and a persulfidated cysteine on the candidate substrate protein in cells. Largely non-overlapping sets of four, 32 and three candidate cellular substrates were detected for TrxA, TrxP, and TrxQ, respectively, many of which were previously identified as global proteome S-sulfuration targets including for example, pyruvate kinase, PykA. Both TrxA (kcat = 0.13 s-1) and TrxP (kcat = 0.088 s-1) are capable of reducing protein persulfides on PykA, a model substrate detected as a candidate substrate of TrxP; in contrast, TrxQ shows lower activity (kcat = 0.015 s-1). This work reveals that protein S-sulfuration, central to H2S and reactive sulfur species (RSS) signaling, may impact cellular activities and appears to be regulated in S. aureus largely by TrxP under conditions of sulfide stress.https://www.frontiersin.org/article/10.3389/fmicb.2018.02385/fullthioredoxin-like proteinthioredoxin profilinghydrogen sulfideprotein persulfide reductionpyruvate kinase |
| spellingShingle | Hui Peng Hui Peng Yixiang Zhang Yixiang Zhang Jonathan C. Trinidad Jonathan C. Trinidad David P. Giedroc David P. Giedroc Thioredoxin Profiling of Multiple Thioredoxin-Like Proteins in Staphylococcus aureus Frontiers in Microbiology thioredoxin-like protein thioredoxin profiling hydrogen sulfide protein persulfide reduction pyruvate kinase |
| title | Thioredoxin Profiling of Multiple Thioredoxin-Like Proteins in Staphylococcus aureus |
| title_full | Thioredoxin Profiling of Multiple Thioredoxin-Like Proteins in Staphylococcus aureus |
| title_fullStr | Thioredoxin Profiling of Multiple Thioredoxin-Like Proteins in Staphylococcus aureus |
| title_full_unstemmed | Thioredoxin Profiling of Multiple Thioredoxin-Like Proteins in Staphylococcus aureus |
| title_short | Thioredoxin Profiling of Multiple Thioredoxin-Like Proteins in Staphylococcus aureus |
| title_sort | thioredoxin profiling of multiple thioredoxin like proteins in staphylococcus aureus |
| topic | thioredoxin-like protein thioredoxin profiling hydrogen sulfide protein persulfide reduction pyruvate kinase |
| url | https://www.frontiersin.org/article/10.3389/fmicb.2018.02385/full |
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