Advertisement
JBC

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Originally published In Press as doi:10.1074/jbc.C700105200 on July 4, 2007

J. Biol. Chem., Vol. 282, Issue 36, 25981-25985, September 7, 2007
This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Supplemental Data
Right arrow All Versions of this Article:
282/36/25981    most recent
C700105200v1
Right arrow Submit a Letter to Editor
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowRequest Permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Hochgräfe, F.
Right arrow Articles by Hecker, M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Hochgräfe, F.
Right arrow Articles by Hecker, M.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

S-Cysteinylation Is a General Mechanism for Thiol Protection of Bacillus subtilis Proteins after Oxidative Stress*Formula

Falko Hochgräfe{ddagger}, Jörg Mostertz{ddagger}1, Dierk-Christoph Pöther{ddagger}, Dörte Becher{ddagger}, John D. Helmann§, and Michael Hecker{ddagger}2

From the {ddagger}Institute for Microbiology, Ernst-Moritz-Arndt-University Greifswald, D-17487 Greifswald, Germany and the §Department of Microbiology, Cornell University, Ithaca, New York 14853-8101

Received for publication, May 30, 2007 , and in revised form, July 2, 2007.


   ABSTRACT
TOP
 ABSTRACT
INTRODUCTION
 EXPERIMENTAL PROCEDURES
 RESULTS AND DISCUSSION
 REFERENCES
 
S-Thiolation is crucial for protection and regulation of thiol-containing proteins during oxidative stress and is frequently achieved by the formation of mixed disulfides with glutathione. However, many Gram-positive bacteria including Bacillus subtilis lack the low molecular weight (LMW) thiol glutathione. Here we provide evidence that S-thiolation by the LMW thiol cysteine represents a general mechanism in B. subtilis. In vivo labeling of proteins with [35S]cysteine and nonreducing two-dimensional PAGE analyses revealed that a large subset of proteins previously identified as having redox-sensitive thiols are modified by cysteine in response to treatment with the thiol-specific oxidant diamide. By means of multidimensional shotgun proteomics, the sites of S-cysteinylation for six proteins could be identified, three of which are known to be S-glutathionylated in other organisms.


   INTRODUCTION
TOP
 ABSTRACT
 INTRODUCTION
EXPERIMENTAL PROCEDURES
 RESULTS AND DISCUSSION
 REFERENCES
 
Protein S-thiolation by low molecular weight (LMW)3 thiols prevents the irreversible oxidation of cysteine residues during oxidative stress and plays a pivotal role in the redox regulation of thiol-containing proteins. For example, the formation of mixed disulfides between target proteins and the LMW thiol glutathione is a key event in the regulation of the eukaryotic proteins, including c-Jun, thioredoxin, and glyceraldehyde-3-phosphate dehydrogenase (1–5). S-Glutathionylation has also been reported to modulate the activity of Escherichia coli methionine synthase (MetE), phosphoadenosine phosphosulfate reductase (CysH), and the oxidative stress-specific transcription factor OxyR (6–8).

Because many Gram-positive bacteria lack glutathione, the nature of S-thiolation in these organisms remains elusive (9). In the Gram-positive model organism Bacillus subtilis, cysteine represents the most abundant LMW thiol (9). One of the most obvious responses of B. subtilis to disulfide stress is the strong induction of cysteine biosynthesis genes (10). Although the origins of this effect are unclear, it might reflect a consumption of free cysteine by oxidation to cystine and the formation of mixed disulfides with proteins.

In a previous proteome analysis, we reported that reversible thiol oxidation occurs in a number of B. subtilis proteins after oxidative stress indicating a general thiol-protection mechanism in this organism (11). We also showed that overoxidation of cysteine residues to sulfonic acid caused by high level peroxide stress results in protein damage and in the irreversible loss of protein activity in B. subtilis and Staphylococcus aureus (11, 12). However, the nature of the reversible thiol modifications was not apparent and likely reflected a mixture of intra- or intermolecular protein disulfides as well as mixed disulfides with LMW thiols. Recently, Lee et al. (13) demonstrated that the organic peroxide-sensing repressor OhrR of B. subtilis is reversibly inactivated by S-thiolation by cysteine, an unknown thiol of 398 Da, or coenzyme A (CoASH) under conditions of oxidative stress. S-Thiolation by CoASH also has been shown to occur during spore formation in Bacillus megaterium and 45% of the CoASH in spores was found as mixed disulfides with protein (14).

In the present study, we conducted a global survey to detect proteins modified by S-cysteinylation under conditions of disulfide stress. By in vivo [35S]cysteine labeling and two-dimensional PAGE, a large subset of proteins was observed, which showed S-thiolation in response to diamide treatment. Multidimensional "shotgun" proteomics revealed S-cysteinylation and identified the sites of this modification in six different B. subtilis proteins. No evidence was obtained under these conditions for S-thiolation by either CoASH or the recently described 398-Da LMW thiol (13, 15). Our results suggest that S-cysteinylation represents a general mechanism of thiol protection and functions as an actual mode of redox regulation under disulfide stress conditions in B. subtilis.


   EXPERIMENTAL PROCEDURES
TOP
 ABSTRACT
 INTRODUCTION
 EXPERIMENTAL PROCEDURES
RESULTS AND DISCUSSION
 REFERENCES
 
Strain and Growth Medium—The B. subtilis wild type strain 168 (16) was grown aerobically at 37 °C in a synthetic medium (17).

[35S]Cysteine Monitoring of S-Thiolations—To prevent protein synthesis, 500µg/ml chloramphenicol was added to exponentially growing cells (A500 nm 0.5) 30 min before cell treatment. The cells were incubated with 10 µCi/ml of L-[35S]cysteine (specific activity 1,000 Ci/mmol and radioactive concentration 10 mCi/ml at reference date, GE Healthcare) for 20 min to provide labeled cysteine. Conditions of oxidative stress were achieved by the addition of a final concentration of 1 mM diamide (11, 18) for 30 min. For control purposes, cultivation was carried out without the addition of diamide.

Cell Sampling and Blocking of Thiol Groups—Cells were harvested by centrifugation with 8,900 x g for 5 min at 4 °C. Subsequently, the resulting pellet was resuspended in denaturing buffer (8 M urea, 1% CHAPS, 1 mM EDTA, 200 mM Tris-HCl, pH 8.0) containing 100 mM iodoacetamide. Crude cell extracts were obtained by sonication and were then incubated for 20 min at room temperature to alkylate accessible free thiol groups. To terminate the blocking reaction, proteins were precipitated by addition of 4 parts ice-cold acetone and stored at –20 °C for 1 h. Samples were then centrifuged at 21,900 x g for 15 min at 20 °C and washed with 80% acetone and pure acetone. The resulting protein pellet was dried in a vacuum centrifuge. Samples were dissolved in 150 µl of denaturing buffer without iodoacetamide and the protein concentration was determined. To prove reversible S-thiolations, aliquots of the labeled protein extracts were treated with the disulfide reducing agent at final concentrations of 10 mM dithiothreitol or 1 mM Tris-(2-carboxyethyl)-phosphine for 30 min at room temperature.

Measurement of Protein-associated Radioactivity with and without Reducing Agents—Similar protein amounts (150 and 300 µg) of the untreated or reduced samples were loaded on filter disks. The protein bound radioactivity was measured in a scintillation counter (19). A dilution series of L-[35S]cysteine from 2 to 1000 nCi was used to calculate the radioactivity from the measured counts per minute.

Nonreducing Two-dimensional PAGE—Similar protein amounts (100 µg) with 35S-labeled S-thiolations were separated by two-dimensional PAGE according to Büttner et al. (20). To ensure nonreducing conditions dithiothreitol and thiourea were omitted from the procedure. Gels were stained with silver nitrate, and total protein images were obtained using a light scanner with a transparency unit. For autoradiograms, gels were dried on filter paper, exposed to storage phosphor screens, and scanned with the Typhoon 9400 variable mode imager (Amersham Biosciences).

Shotgun Proteomics—Diamide was added to exponentially growing cells (A500 nm 0.5) to a final concentration of 1 mM for 20 min. Cultivation was carried out without the addition of diamide for control purposes. Cells were harvested, and thiol groups were blocked as described above. The protein mixture was mixed with nonreducing PAGE loading buffer and separated by electrophoresis in a one-dimensional SDS-PAGE mini-gel. In-gel digestion of equal gel pieces was carried out with trypsin as described recently (21). The peptide mixture was subjected to a reversed phase separation using the Ettan MDLC System (GE Healthcare) and analyzed using a LTQ FTICR mass spectrometer (Thermo Electron Corp., San Jose, CA) (22). MS data sets were analyzed as described in the legend for supplemental Fig. S2. All data were obtained from two independently performed MS analyses of at least two independent cell cultivation experiments.


   RESULTS AND DISCUSSION
TOP
 ABSTRACT
 INTRODUCTION
 EXPERIMENTAL PROCEDURES
 RESULTS AND DISCUSSION
REFERENCES
 
To find out whether B. subtilis uses free cysteine to form mixed protein disulfides under conditions of disulfide stress, we analyzed cells treated with the disulfide-generating agent diamide by means of the [35S]cysteine labeling approach (23). In brief, growing cells were treated with chloramphenicol to prevent protein synthesis followed by the addition of L-[35S]cysteine to provide free labeled cysteine. To provoke thiol oxidation, the cells were stressed with diamide (11, 18). After extraction of intracellular proteins, the protein-associated radioactivity was finally measured. Compared with untreated control cells, intracellular proteins of cells treated with diamide showed an approximately 6-fold increase in the amount of 35S radioactivity (Fig. 1).


Figure 1
View larger version (15K):
[in this window]
[in a new window]

  		FIGURE 1.
Monitoring of protein S-thiolations. Cells of B. subtilis were supplemented with [35S]cysteine, and protein synthesis was prevented by chloramphenicol. Diamide was added to provoke thiol oxidation. The 35S signal of the protein-associated radioactivity is shown as fmol of [35S]cysteine incorporation/mg of soluble protein (gray bars). For determination of the extent of S-thiolations, the labeled protein extracts were treated with disulfide-reducing agents (white bars). The values are given as means ± S.D. of three parallel studies of three independently analyzed cell cultures.

 
To evaluate whether the detected increase in 35S-protein labeling is a reversible thiol modification, we incubated aliquots of the labeled protein extracts with disulfide reducing agents. The reduction step resulted in about an 81% loss of radioactivity in the diamide-treated sample, indicating that the vast majority of 35S labeling accounts for mixed protein disulfides initiated by cysteine (Fig. 1).

On the basis of these results, we considered that calculating the amount of protein bound [35S]cysteine could give us a lower limit of the number of S-thiolations per cell. We estimated about a 30-fold increase in cysteine-based S-thiolation events per cell after 30 min of diamide treatment. This corresponds to an absolute number of at least 1000 cysteine-based S-thiolation events in one single cell after disulfide stress. Because of putative S-thiolation using unlabeled cysteine and significant losses of thiol modifications during the experimental procedure itself, we have provided this number as the lower limit.


Figure 2
View larger version (35K):
[in this window]
[in a new window]

  		FIGURE 2.
Mass spectrometric identification of the S-cysteinylation at Cys-719 in the cobalamin-independent methionine synthase MetE from cell extracts of diamide treated B. subtilis cells. The tryptic peptide containing amino acid residues Val-702 to Arg-723 modified by the additional S-cysteinylation-specific mass of 119 Da was assigned to the detected precursor ion of 852.4 [M+3H]3+ and the corresponding fragment ion spectrum. Mass peaks that match the expected b- and y-ions are highlighted.

 
To visualize the overall pattern of S-thiolated proteins, [35S]cysteine-labeled protein extracts were separated by two-dimensional gel electrophoresis (20) under nonreducing conditions. The autoradiograms of the two-dimensional gels revealed a strong increase in the number of protein spots with associated 35S radioactivity after diamide stress, again indicating S-thiolation after oxidative stress (see supplemental Fig. S1). These results suggest that many intracellular proteins of B. subtilis are readily S-thiolated by free cysteine (or a derivative) after diamide treatment.

To characterize the precise molecular nature of S-thiolations and identify the specific sites of the S-thiolation events, we surveyed cytoplasmic proteins from diamide-treated cells by multidimensional shotgun proteomics using a LTQ FTICR mass spectrometer. In summary, growing cells were treated with diamide to provoke severe reversible thiol oxidation. The protein fraction was separated by nonreducing one-dimensional protein gel electrophoresis, and in-gel digestion by trypsin using gel pieces was carried out. Peptide mixtures were then subjected to a reversed phase separation using a nanoscale liquid chromatography system and were measured with the LTQ FTICR mass spectrometer. The data were analyzed using the SEQUEST algorithm and a data base of B. subtilis proteins as described in the legend for supplemental Fig. S2. In brief, the data set was scanned for peptides with mixed disulfide bonds to cysteine, homocysteine, N-acetylcysteine, CoASH, {gamma}-glutamylcysteine, glutathione, or the novel 398-Da LMW thiol. The filtering was achieved by using BioworksBrowser software to identify peptides with a mass deviation below 2 ppm between the measured and the calculated mass (see supplemental Fig. S2 for details).

The LTQ FTICR analysis revealed sites of S-cysteinylation in six different peptides from six different cysteine-containing proteins after disulfide stress (Table 1). The modified proteins included inosine-monophosphate dehydrogenase (GuaB), cobalamin-independent methionine synthase (MetE), and inorganic pyrophosphatase (PpaC). All six proteins showed an additional mass of 119 Da at a cysteine-containing peptide, which is specific for S-cysteinylation (Fig. 2 and supplemental Fig. S2). The fragment ion spectra of the target peptides were used to confirm the exact position of the S-cysteinylation at the cysteine within the amino acid sequence of the corresponding peptide (Fig. 2 and Table 1). In peptide mixtures derived from control cells (grown without diamide) no S-thiolated proteins were detected, indicating that protein S-cysteinylation accumulates during oxidative stress.


View this table:
[in this window]
[in a new window]

  		TABLE 1
Proteins with S-cysteinylations after diamide stress in B. subtilis

 
Four of the six identified S-cysteinylated proteins, namely GuaB, MetE, PpaC, and the protein YwaA with similarity to branched-chain amino acid aminotransferases, were already reported to show increased amounts of reversible thiol oxidation after oxidative stress using a fluorescence-based thiol modification assay (11). Three homologous proteins (GuaB, MetE, and PpaC) are known to be S-glutathionylated in other organisms (6, 24, 25) (Table 1).

The inosine-monophosphate dehydrogenase GuaB was found to be S-cysteinylated at its active site cysteine (Cys-308). A regulatory significance of this GuaB S-thiolation is yet unknown, but our results point to a S-cysteinylation-dependent inhibition of GuaB, which in turn may cause an arrest of GTP biosynthesis and therefore may help to prevent the accumulation of the highly mutagenic 7,8-dihydro-8-oxoguanine (26) under conditions of oxidative stress.

For E. coli, it has been shown that the inhibition of methionine biosynthesis is caused by the reversible thiol oxidation of methionine synthase MetE after oxidative stress (6). Likewise, we have demonstrated thiol oxidation of MetE and methionine auxotrophy in B. subtilis cells after diamide or peroxide treatment (11). In contrast to the S-glutathionylation that was detected for MetE of E. coli, our present study shows that MetE of B. subtilis is S-cysteinylated on Cys-719. We therefore suggest that B. subtilis MetE is protected and reversibly inactivated by S-cysteinylation at Cys-719 under conditions of oxidative stress. In B. subtilis, the cysteine biosynthesis is tightly controlled by different regulators, including CymR, CysL, and Spx (27–30). In response to disulfide stress, B. subtilis cells initiate a strong induction of cysteine biosynthesis genes including the methionine to cysteine conversion pathway (10, 31). The activation of cysteine biosynthesis after thiol-specific oxidative stress is supposed to be caused by depletion of the intracellular cysteine pool following highly elevated S-cysteinylation to protect redox-sensitive thiols from irreversible oxidation. Therefore, it can be presumed that the reported inhibitory S-cysteinylation of MetE at Cys-719 contributes in concert with the strong induction of cysteine biosynthesis genes to redirecting the metabolism from methionine toward cysteine biosynthesis for LMW thiol generation.

A high number of proteins were detected to be S-thiolated by the [35S]cysteine assay. The six proteins identified with S-cysteinylation by mass spectrometry appear to be just the tip of the iceberg, because in this initial study we obtained only proteins with S-cysteinylation that were reported to be among the 115 most abundant proteins in growing B. subtilis cells (21). Apart from the S-cysteinylation, no other type of S-thiolation could be detected in B. subtilis cells after diamide treatment. The lack of S-glutathionylation and S-{gamma}-glutamylcysteinylation may be explained by the lack of the genes encoding the corresponding enzymes for biosynthesis of glutathione or {gamma}-glutamylcysteine in B. subtilis. The failure to detect mixed disulfides with CoASH and the recently described 398-Da LMW thiol (13, 32) may be due to a comparatively low abundance of these thiols, a preferential reaction of diamide with cysteine versus other LMW thiols, or a differential sensitivity of detection of the various modified peptides by LTQ FTICR mass spectrometry. It is known, for example, that CoASH-modified peptides are very poorly detected in positive ion mode mass spectrometry studies, likely because of the poor ionization of the phosphate-containing CoASH moiety (32). Further analyses will be required to develop methods to accurately quantify the various LMW thiols in B. subtilis, and their relative contributions to thiol modification under various oxidative stress conditions.

In the present work, we have demonstrated that the S-thiolation of proteins of B. subtilis can be detected by the addition of labeled free cysteine during oxidative stress treatment. Importantly, we were able to identify six different proteins by multidimensional shotgun proteomics that showed in vivo S-cysteinylation after disulfide stress. Taken together, our results suggest that S-cysteinylation is a general mechanism to protect protein thiols from further oxidation and irreversible damage in B. subtilis and may play a role in redox regulation of several thiol-containing proteins.


   FOOTNOTES
 
* This work was supported by grants from the European Union (LSHC-CT-2004-503468), the Deutsche Forschungsgemeinschaft (HE 1887/7-4), and the Bildungsministerium of des Landes Mecklenburg-Vorpommern (to M. H.) and by NSF Grant MCB-0640616 (to J. D. H.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. Back

Formula The on-line version of this article (available at http://www.jbc.org) contains supplemental Figs. S1 and S2. Back

1 Present address: Interfaculty Inst. for Genetics and Functional Genomics, Junior Research Group Transcriptomics, University of Greifswald, 17489 Greifswald, Germany. Back

2 To whom correspondence should be addressed. Tel.: 49-3834-864200; Fax: 49-3834-864202; E-mail: hecker{at}uni-greifswald.de.

3 The abbreviations used are: LMW, low molecular weight; MS, mass spectrometry; LTQ FTICR, linear ion trap Fourier transform ion cyclotron resonance. Back


   ACKNOWLEDGMENTS
 
We thank Antje Plotz for excellent technical assistance.



   REFERENCES
TOP
 ABSTRACT
 INTRODUCTION
 EXPERIMENTAL PROCEDURES
 RESULTS AND DISCUSSION
 REFERENCES
 

  1. Klatt, P., Molina, E. P., and Lamas, S. (1999) J. Biol. Chem. 274, 15857–15864[Abstract/Free Full Text]
  2. Klatt, P., Molina, E. P., De Lacoba, M. G., Padilla, C. A., Martinez-Galesteo, E., Barcena, J. A., and Lamas, S. (1999) FASEB J. 13, 1481–1490[Abstract/Free Full Text]
  3. Casagrande, S., Bonetto, V., Fratelli, M., Gianazza, E., Eberini, I., Massignan, T., Salmona, M., Chang, G., Holmgren, A., and Ghezzi, P. (2002) Proc. Natl. Acad. Sci. U. S. A. 99, 9745–9749[Abstract/Free Full Text]
  4. Michelet, L., Zaffagnini, M., Marchand, C., Collin, V., Decottignies, P., Tsan, P., Lancelin, J. M., Trost, P., Miginiac-Maslow, M., Noctor, G., and Lemaire, S. D. (2005) Proc. Natl. Acad. Sci. U. S. A. 102, 16478–16483[Abstract/Free Full Text]
  5. Cotgreave, I. A., Gerdes, R., Schuppe-Koistinen, I., and Lind, C. (2002) Methods Enzymol. 348, 175–182[Medline] [Order article via Infotrieve]
  6. Hondorp, E. R., and Matthews, R. G. (2004) PLoS Biol. 2, e336[CrossRef][Medline] [Order article via Infotrieve]
  7. Lillig, C. H., Potamitou, A., Schwenn, J. D., Vlamis-Gardikas, A., and Holmgren, A. (2003) J. Biol. Chem. 278, 22325–22330[Abstract/Free Full Text]
  8. Kim, S. O., Merchant, K., Nudelman, R., Beyer, W. F., Jr., Keng, T., DeAngelo, J., Hausladen, A., and Stamler, J. S. (2002) Cell 109, 383–396[CrossRef][Medline] [Order article via Infotrieve]
  9. Newton, G. L., Arnold, K., Price, M. S., Sherrill, C., Delcardayre, S. B., Aharonowitz, Y., Cohen, G., Davies, J., Fahey, R. C., and Davis, C. (1996) J. Bacteriol. 178, 1990–1995[Abstract/Free Full Text]
  10. Leichert, L. I., Scharf, C., and Hecker, M. (2003) J. Bacteriol. 185, 1967–1975[Abstract/Free Full Text]
  11. Hochgräfe, F., Mostertz, J., Albrecht, D., and Hecker, M. (2005) Mol. Microbiol. 58, 409–425[CrossRef][Medline] [Order article via Infotrieve]
  12. Weber, H., Engelmann, S., Becher, D., and Hecker, M. (2004) Mol. Microbiol. 52, 133–140[CrossRef][Medline] [Order article via Infotrieve]
  13. Lee, J. W., Soonsanga, S., and Helmann, J. D. (2007) Proc. Natl. Acad. Sci. U. S. A. 104, 8743–8748[Abstract/Free Full Text]
  14. Setlow, B., and Setlow, P. (1977) Biochem. Biophys. Res. Commun. 75, 45–50[CrossRef][Medline] [Order article via Infotrieve]
  15. Nicely, N. I., Parsonage, D., Paige, C., Newton, G. L., Fahey, R. C., Leonardi, R., Jackowski, S., Mallett, T. C., and Claiborne, A. (2007) Biochemistry 46, 3234–3245[CrossRef][Medline] [Order article via Infotrieve]
  16. Anagnostopoulos, C., and Spizizen, J. (1961) J. Bacteriol. 81, 741–746[Free Full Text]
  17. Stülke, J., Hanschke, R., and Hecker, M. (1993) J. Gen. Microbiol. 139, 2041–2045[Medline] [Order article via Infotrieve]
  18. Kosower, N. S., and Kosower, E. M. (1995) Methods Enzymol. 251, 123–133[Medline] [Order article via Infotrieve]
  19. Bandow, J. E., Brötz, H., and Hecker, M. (2002) J. Bacteriol. 184, 459–467[Abstract/Free Full Text]
  20. Büttner, K., Bernhardt, J., Scharf, C., Schmid, R., Mäder, U., Eymann, C., Antelmann, H., Völker, A., Völker, U., and Hecker, M. (2001) Electrophoresis 22, 2908–2935[CrossRef][Medline] [Order article via Infotrieve]
  21. Eymann, C., Dreisbach, A., Albrecht, D., Bernhardt, J., Becher, D., Gentner, S., Tam, le T., Büttner, K., Buurman, G., Scharf, C., Venz, S., Völker, U., and Hecker, M. (2004) Proteomics 4, 2849–2876[CrossRef][Medline] [Order article via Infotrieve]
  22. Wolff, S., Otto, A., Albrecht, D., Zeng, J. S., Büttner, K., Glückmann, M., Hecker, M., and Becher, D. (2006) Mol. Cell. Proteomics 5, 1183–1192[Abstract/Free Full Text]
  23. Ghezzi, P., Romines, B., Fratelli, M., Eberini, I., Gianazza, E., Casagrande, S., Laragione, T., Mengozzi, M., and Herzenberg, L. A. (2002) Mol. Immunol. 38, 773–780[CrossRef][Medline] [Order article via Infotrieve]
  24. Lind, C., Gerdes, R., Hamnell, Y., Schuppe-Koistinen, I., von Lowenhielm, H. B., Holmgren, A., and Cotgreave, I. A. (2002) Arch. Biochem. Biophys. 406, 229–240[CrossRef][Medline] [Order article via Infotrieve]
  25. Fratelli, M., Gianazza, E., and Ghezzi, P. (2004) Expert Rev. Proteomics 1, 365–376[CrossRef][Medline] [Order article via Infotrieve]
  26. Henle, E. S., and Linn, S. (1997) J. Biol. Chem. 272, 19095–19098[Free Full Text]
  27. Erwin, K. N., Nakano, S., and Zuber, P. (2005) J. Bacteriol. 187, 4042–4049[Abstract/Free Full Text]
  28. Even, S., Burguiere, P., Auger, S., Soutourina, O., Danchin, A., and Martin-Verstraete, I. (2006) J. Bacteriol. 188, 2184–2197[Abstract/Free Full Text]
  29. Choi, S. Y., Reyes, D., Leelakriangsak, M., and Zuber, P. (2006) J. Bacteriol. 188, 5741–5751[Abstract/Free Full Text]
  30. Guillouard, I., Auger, S., Hullo, M. F., Chetouani, F., Danchin, A., and Martin-Verstraete, I. (2002) J. Bacteriol. 184, 4681–4689[Abstract/Free Full Text]
  31. Hullo, M. F., Auger, S., Soutourina, O., Barzu, O., Yvon, M., Danchin, A., and Martin-Verstraete, I. (2007) J. Bacteriol. 189, 187–197[Abstract/Free Full Text]
  32. Janek, K., Wenschuh, H., Bienert, M., and Krause, E. (2001) Rapid Commun. Mass Spectrom. 15, 1593–1599[CrossRef][Medline] [Order article via Infotrieve]
  33. Hao, G., Xie, L., and Gross, S. S. (2004) J. Biol. Chem. 279, 36192–36200[Abstract/Free Full Text]
  34. Lahti, R., and Suonpaa, M. (1982) J. Gen. Microbiol. 128, 1023–1026[Abstract/Free Full Text]

Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?


This article has been cited by other articles:


Home page
 MicrobiologyHome page
V. Barbe, S. Cruveiller, F. Kunst, P. Lenoble, G. Meurice, A. Sekowska, D. Vallenet, T. Wang, I. Moszer, C. Medigue, et al.
From a consortium sequence to a unified sequence: the Bacillus subtilis 168 reference genome a decade later
Microbiology, June 1, 2009; 155(6): 1758 - 1775.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
E. R. Hondorp and R. G. Matthews
Oxidation of Cysteine 645 of Cobalamin-Independent Methionine Synthase Causes a Methionine Limitation in Escherichia coli
J. Bacteriol., May 15, 2009; 191(10): 3407 - 3410.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
R. Lo, M. S. Turner, D. G. Barry, R. Sreekumar, T. P. Walsh, and P. M. Giffard
Cystathionine {gamma}-Lyase Is a Component of Cystine-Mediated Oxidative Defense in Lactobacillus reuteri BR11
J. Bacteriol., March 15, 2009; 191(6): 1827 - 1837.
[Abstract] [Full Text] [PDF]

Home page
 Mol PlantHome page
X.-H. Gao, M. Bedhomme, D. Veyel, M. Zaffagnini, and S. D. Lemaire
Methods for Analysis of Protein Glutathionylation and their Application to Photosynthetic Organisms
Mol Plant, March 1, 2009; 2(2): 218 - 235.
[Abstract] [Full Text] [PDF]

Home page
 Appl. Environ. Microbiol.Home page
T. R. H. M. Kouwen, J.-Y. F. Dubois, R. Freudl, W. J. Quax, and J. M. van Dijl
Modulation of Thiol-Disulfide Oxidoreductases for Increased Production of Disulfide-Bond-Containing Proteins in Bacillus subtilis
Appl. Envir. Microbiol., December 15, 2008; 74(24): 7536 - 7545.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
C. Paige, S. D. Reid, P. C. Hanna, and A. Claiborne
The Type III Pantothenate Kinase Encoded by coaX Is Essential for Growth of Bacillus anthracis
J. Bacteriol., September 15, 2008; 190(18): 6271 - 6275.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
K. Helbig, C. Bleuel, G. J. Krauss, and D. H. Nies
Glutathione and Transition-Metal Homeostasis in Escherichia coli
J. Bacteriol., August 1, 2008; 190(15): 5431 - 5438.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
K. Helbig, C. Grosse, and D. H. Nies
Cadmium Toxicity in Glutathione Mutants of Escherichia coli
J. Bacteriol., August 1, 2008; 190(15): 5439 - 5454.
[Abstract] [Full Text] [PDF]

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Supplemental Data
Right arrow All Versions of this Article:
282/36/25981    most recent
C700105200v1
Right arrow Submit a Letter to Editor
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowRequest Permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Hochgräfe, F.
Right arrow Articles by Hecker, M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Hochgräfe, F.
Right arrow Articles by Hecker, M.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
 All ASBMB Journals   Molecular and Cellular Proteomics 
 Journal of Lipid Research   ASBMB Today 
Copyright © 2007 by the American Society for Biochemistry and Molecular Biology.
Advertisement
spacer
Advertisement
Advertisement