Prototype of a heme chaperone essential for cytochrome c maturation

Science

Volumn 281, Issue 5380, 1998, Pages 1197-1200

  Schulz, Henk ^a   Hennecke, Hauke ^a   Thöny Meyer, Linda ^a  

^a ETH ZURICH   (Switzerland)

Author keywords

[No Author keywords available]

Indexed keywords

APOENZYME; CHAPERONE; CYTOCHROME C; HEME; HEMOPROTEIN; HISTIDINE;

ARTICLE; COMPLEX FORMATION; CONTROLLED STUDY; COVALENT BOND; ESCHERICHIA COLI; MATURATION; NONHUMAN; PRIORITY JOURNAL; PROTEIN ANALYSIS;

BACTERIA (MICROORGANISMS); ESCHERICHIA COLI; NEGIBACTERIA;

EID: 0032555539     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.281.5380.1197     Document Type: Article

References (34)

1. L. Thöny-Meyer, D. Ritz, H. Hennecke, Mol. Microbiol. 12, 1 (1994); G. Howe and S. Merchant, Photosynth. Res. 40, 147 (1994); M. D. Page et al., Trends Biochem. Sci. 23, 103 (1998); R. G. Kranz et al., Mol. Microbiol. 29, 383 (1998).
2. L. Thöny-Meyer, D. Ritz, H. Hennecke, Mol. Microbiol. 12, 1 (1994); G. Howe and S. Merchant, Photosynth. Res. 40, 147 (1994); M. D. Page et al., Trends Biochem. Sci. 23, 103 (1998); R. G. Kranz et al., Mol. Microbiol. 29, 383 (1998).
3. L. Thöny-Meyer, D. Ritz, H. Hennecke, Mol. Microbiol. 12, 1 (1994); G. Howe and S. Merchant, Photosynth. Res. 40, 147 (1994); M. D. Page et al., Trends Biochem. Sci. 23, 103 (1998); R. G. Kranz et al., Mol. Microbiol. 29, 383 (1998).
4. L. Thöny-Meyer, D. Ritz, H. Hennecke, Mol. Microbiol. 12, 1 (1994); G. Howe and S. Merchant, Photosynth. Res. 40, 147 (1994); M. D. Page et al., Trends Biochem. Sci. 23, 103 (1998); R. G. Kranz et al., Mol. Microbiol. 29, 383 (1998).
5. L. Thöny-Meyer, Microbiol. Mol. Biol. Rev. 61, 337 (1997).
6. D. H. Gonzales and W. Neupert, J. Bioenerg. Biomembr. 22, 753 (1990).
7. H. A Dailey, in Biosynthesis of Heme and Chlorophylls (McGraw-Hill, New York, 1990), pp. 123-160.
8. Y. Sambongi, R. Stoll, S. J. Ferguson, Mol. Microbiol. 19, 1193 (1996).
9. L. Thöny-Meyer and P. Künzler, Eur. J. Biochem. 246, 794 (1997).
10. G. Basile, C. Di Bello, H. Taniuchi, J. Biol. Chem. 255, 7181 (1980); B. Hennig and W. Neupert, Eur. J. Biochem. 121, 203 (1981).
11. G. Basile, C. Di Bello, H. Taniuchi, J. Biol. Chem. 255, 7181 (1980); B. Hennig and W. Neupert, Eur. J. Biochem. 121, 203 (1981).
12. L. Thöny-Meyer et al., J. Bacteriol. 177, 4321 (1995).
13. J. Grove et al., Mol. Microbiol. 19, 467 (1996).
14. 94) was constructed by polymerase chain reaction (PCR)-based cloning and subsequent gene replacement mutagenesis of the wild-type strain MC1061. A pMAK705-derived plasmid with a temperature-sensitive origin of replication [C. M. Hamilton et al., J. Bacteriol. 171, 4617 (1989) ] containing the ccmA gene expressed constitutively from the pA-CYC184-derived tet promoter [A. C. Y. Chang and S. N. Cohen, ibid. 134, 1141 (1978)] was cointegrated into the wild type at high temperatures to obtain enhanced and anaerobiosis-independent expression of the ccm genes, which stimulated holocytochrome c formation. An equivalent strain carrying the ΔccmE mutation (ΔccmE cointegrate) was obtained by cointegrating the same plasmid into the chromosomal ΔccmE mutant. The ΔccmE cointegrate was used as a background for expression of the various CcmE proteins described in this work.
15. 94) was constructed by polymerase chain reaction (PCR)-based cloning and subsequent gene replacement mutagenesis of the wild-type strain MC1061. A pMAK705-derived plasmid with a temperature-sensitive origin of replication [C. M. Hamilton et al., J. Bacteriol. 171, 4617 (1989) ] containing the ccmA gene expressed constitutively from the pA-CYC184-derived tet promoter [A. C. Y. Chang and S. N. Cohen, ibid. 134, 1141 (1978)] was cointegrated into the wild type at high temperatures to obtain enhanced and anaerobiosis-independent expression of the ccm genes, which stimulated holocytochrome c formation. An equivalent strain carrying the ΔccmE mutation (ΔccmE cointegrate) was obtained by cointegrating the same plasmid into the chromosomal ΔccmE mutant. The ΔccmE cointegrate was used as a background for expression of the various CcmE proteins described in this work.
16. 94) was constructed by polymerase chain reaction (PCR)-based cloning and subsequent gene replacement mutagenesis of the wild-type strain MC1061. A pMAK705-derived plasmid with a temperature-sensitive origin of replication [C. M. Hamilton et al., J. Bacteriol. 171, 4617 (1989) ] containing the ccmA gene expressed constitutively from the pA-CYC184-derived tet promoter [A. C. Y. Chang and S. N. Cohen, ibid. 134, 1141 (1978)] was cointegrated into the wild type at high temperatures to obtain enhanced and anaerobiosis-independent expression of the ccm genes, which stimulated holocytochrome c formation. An equivalent strain carrying the ΔccmE mutation (ΔccmE cointegrate) was obtained by cointegrating the same plasmid into the chromosomal ΔccmE mutant. The ΔccmE cointegrate was used as a background for expression of the various CcmE proteins described in this work.
17. ara in the presence of arabinose.
18. 14C]-δ-aminolevulinic acid (1.762 GBq/mmol, NEN) was added. After 1 hour, proteins were precipitated with TCA and applied to 15% SOS-PAGE. Radioactively labeled hemoproteins were detected on a Phosphorlmager SF (Molecular Dynamics).
19. 2+ affinity chromatography on nitrilotriacetic acid (NTA) agarose (Quiagen).
20. -1 in the pyridine hemochrome assay. CcmE polypeptide concentration was quantified after S. C. Gill and H. P. van Hippel [Anal. Biochem. 182, 319 (1989)].
21. -1 in the pyridine hemochrome assay. CcmE polypeptide concentration was quantified after S. C. Gill and H. P. van Hippel [Anal. Biochem. 182, 319 (1989)].
22. C. Manoil and J. Beckwith, Science 233, 1403 (1986).
23. ara promoter for overproduction. Both CcmE versions were expressed in a ΔccmE background upon induction with arabinose.
24. 6 was digested with trypsin (1% w/w) at 37°C for 5 hours, and the peptides were separated on a LiChrospher RP-8 HPLC-Cartridge (Hewlett-Packard) with a linear acetonitrile gradient in 0.1% (v/v) TFA. Peptides were monitored at 215 and 415 nm to detect the heme-binding peptide. The peak fraction was collected, lyophilized, and resuspended in 50% (v/v) acetonitrile, 0.1% (v/v) acetate.
25. Abbreviations for the amino acid residues are as follows: A, Ala; C, Cys; D, Asp; E, Glu; F, Phe; G, Gly; H, His; I, Ile; K, Lys; L, Leu; M, Met; N, Asn; P, Pro; Q, Gln; R, Arg; S, Ser; T, Thr; V, Val; W, Trp; and Y, Tyr.
26. MS/MS analysis was performed with a micro-ion-spray ion source on a API 365 LC/MS/MS system (Perkin-Elmer) with a flow rate of 15 μl per hour. The first and third quadrupole were set to scan the mass range of 500 to 2500 m/z. The collision gas in the second quadrupole was nitrogen.
27. L. Thöny-Meyer, P. Künzler, H. Hennecke, Eur. J. Biochem. 235, 754 (1996).
28. F. Grellet et al., GenBank accession number U72502 (1996).
29. B. S. Goldman and R. G Kranz, Mol. Microbiol. 27, 871 (1998).
30. 144 was purchased from TANA Laboratories (Houston, TX).
31. 144 was purchased from TANA Laboratories (Houston, TX).
32. 144 was purchased from TANA Laboratories (Houston, TX).
33. The ccmABCDE genes were expressed constitutively from the pACYC184-derived tet promoter in a Δccm-ABCDEFGH background (8).
34. We thank H. Troxler (Department of Pediatrics, University of Zürich) for mass spectrometry analysis, W. Staudenmann and P. James for protein sequence analysis, A. Hungerbühler for technical assistance, and M. Aebi and A. Helenius for constructive comments on the manuscript. Supported by a grant from the Swiss National Foundation for Scientific Research.