Analysis, by Electrospray Ionization Mass Spectrometry, of Several Forms of Clostridium Pasteurianum Rubredoxin

Overview

Journal Biochem J

Specialty Biochemistry

Date 1993 Dec 15

PMID 8280064

Citations 14

Authors

Y Petillot

E Forest

I Mathieu

J Meyer

J M Moulis

Affiliations

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Abstract

Clostridium pasteurianum rubredoxin and its recombinant counterpart purified from Escherichia coli have been analysed by electrospray ionization m.s. (e.s.i.m.s.). Whereas the N-terminal methionine of the native protein is formylated, the recombinant one has a free N-terminal methionine. E. coli cells also produce a colourless protein from the cloned gene. This protein is absent from C. pasteurianum and was shown to be zinc-substituted rubredoxin. The molecular forms of rubredoxin detected by e.s.i.m.s. depended on the experimental conditions used. Significant conversion into apo-rubredoxin occurred when the proteins were ionized at acidic pH and detected in the positive-ion mode. This conversion was quantitative in the case of Zn-rubredoxin. In contrast, when the proteins were analysed at neutral pH in the negative-ion mode, only the holoproteins, i.e. the species initially present in the solutions, were detected in the spectra. The e.s.i.m.s. experimental conditions set up here may prove useful for the analysis of other acidic metalloproteins with weakly bound metals.

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References

Chait B, Kent S . Weighing naked proteins: practical, high-accuracy mass measurement of peptides and proteins. Science. 1992; 257(5078):1885-94. DOI: 10.1126/science.1411504. View

Hutchens T, Allen M, Li C, Yip T . Occupancy of a C2-C2 type 'zinc-finger' protein domain by copper. Direct observation by electrospray ionization mass spectrometry. FEBS Lett. 1992; 309(2):170-4. DOI: 10.1016/0014-5793(92)81088-4. View

Berg J . Zinc fingers and other metal-binding domains. Elements for interactions between macromolecules. J Biol Chem. 1990; 265(12):6513-6. View

Bergman T, Jornvall H, Holmquist B, Vallee B . A synthetic peptide encompassing the binding site of the second zinc atom (the 'structural' zinc) of alcohol dehydrogenase. Eur J Biochem. 1992; 205(2):467-70. DOI: 10.1111/j.1432-1033.1992.tb16802.x. View

Nar H, Huber R, Messerschmidt A, Filippou A, Barth M, Jaquinod M . Characterization and crystal structure of zinc azurin, a by-product of heterologous expression in Escherichia coli of Pseudomonas aeruginosa copper azurin. Eur J Biochem. 1992; 205(3):1123-9. DOI: 10.1111/j.1432-1033.1992.tb16881.x. View

van de Kamp M, Hali F, Rosato N, Agro A, Canters G . Purification and characterization of a non-reconstitutable azurin, obtained by heterologous expression of the Pseudomonas aeruginosa azu gene in Escherichia coli. Biochim Biophys Acta. 1990; 1019(3):283-92. DOI: 10.1016/0005-2728(90)90206-j. View

Smith A, Santama N, Dacey S, Edwards M, Bray R, Thorneley R . Expression of a synthetic gene for horseradish peroxidase C in Escherichia coli and folding and activation of the recombinant enzyme with Ca2+ and heme. J Biol Chem. 1990; 265(22):13335-43. View

Karlsson B, Pascher T, Nordling M, Arvidsson R, Lundberg L . Expression of the blue copper protein azurin from Pseudomonas aeruginosa in Escherichia coli. FEBS Lett. 1989; 246(1-2):211-7. DOI: 10.1016/0014-5793(89)80285-6. View

Vallee B, Auld D . Zinc coordination, function, and structure of zinc enzymes and other proteins. Biochemistry. 1990; 29(24):5647-59. DOI: 10.1021/bi00476a001. View

10.

Loo J, Loo R, Udseth H, Edmonds C, Smith R . Solvent-induced conformational changes of polypeptides probed by electrospray-ionization mass spectrometry. Rapid Commun Mass Spectrom. 1991; 5(3):101-5. DOI: 10.1002/rcm.1290050303. View

11.

Varadarajan R, Szabo A, Boxer S . Cloning, expression in Escherichia coli, and reconstitution of human myoglobin. Proc Natl Acad Sci U S A. 1985; 82(17):5681-4. PMC: 390615. DOI: 10.1073/pnas.82.17.5681. View

12.

Watenpaugh K, Sieker L, Jensen L . Crystallographic refinement of rubredoxin at 1 x 2 A degrees resolution. J Mol Biol. 1980; 138(3):615-33. DOI: 10.1016/s0022-2836(80)80020-9. View

13.

Eidsness M, ODell S, Kurtz Jr D, Robson R, Scott R . Expression of a synthetic gene coding for the amino acid sequence of Clostridium pasteurianum rubredoxin. Protein Eng. 1992; 5(4):367-71. DOI: 10.1093/protein/5.4.367. View

14.

Omichinski J, Trainor C, Evans T, Gronenborn A, Clore G, Felsenfeld G . A small single-"finger" peptide from the erythroid transcription factor GATA-1 binds specifically to DNA as a zinc or iron complex. Proc Natl Acad Sci U S A. 1993; 90(5):1676-80. PMC: 45942. DOI: 10.1073/pnas.90.5.1676. View

15.

Grupe H, Gottschalk G . Physiological Events in Clostridium acetobutylicum during the Shift from Acidogenesis to Solventogenesis in Continuous Culture and Presentation of a Model for Shift Induction. Appl Environ Microbiol. 1992; 58(12):3896-902. PMC: 183201. DOI: 10.1128/aem.58.12.3896-3902.1992. View

16.

Adams J . On the release of the formyl group from nascent protein. J Mol Biol. 1968; 33(3):571-89. DOI: 10.1016/0022-2836(68)90307-0. View

17.

Meyer J, Gagnon J, Sieker L, Van Dorsselaer A, Moulis J . Rubredoxin from Clostridium thermosaccharolyticum. Amino acid sequence, mass-spectrometric and preliminary crystallographic data. Biochem J. 1990; 271(3):839-41. PMC: 1149642. DOI: 10.1042/bj2710839. View

18.

Mathieu I, Meyer J, Moulis J . Cloning, sequencing and expression in Escherichia coli of the rubredoxin gene from Clostridium pasteurianum. Biochem J. 1992; 285 ( Pt 1):255-62. PMC: 1132774. DOI: 10.1042/bj2850255. View

19.

Vallee B, Coleman J, Auld D . Zinc fingers, zinc clusters, and zinc twists in DNA-binding protein domains. Proc Natl Acad Sci U S A. 1991; 88(3):999-1003. PMC: 50942. DOI: 10.1073/pnas.88.3.999. View

20.

Moulis J, Scherrer N, Gagnon J, Forest E, Petillot Y, Garcia D . Primary structure of Chromatium tepidum high-potential iron-sulfur protein in relation to thermal denaturation. Arch Biochem Biophys. 1993; 305(1):186-92. DOI: 10.1006/abbi.1993.1409. View