Abstract
Ribonucleoside-triphosphate reductase (RTPR, EC 1.17.4.2) from Lactobacillus leichmannii, a monomeric adenosylcobalamin-requiring enzyme, catalyzes the conversion of nucleoside triphosphates to deoxynucleoside triphosphates. The gene for this enzyme has been cloned and sequenced. In contrast to expectations based on mechanistic considerations, there is no statistically significant sequence homology with the Escherichia coli reductase that requires a dinuclear-iron center and tyrosyl radical cofactor. The RTPR has been overexpressed and purified to homogeneity, yielding 90 mg of protein from 2.5 g of bacteria. Initial characterization of the recombinant RTPR indicates that its properties are identical to those of the RTPR isolated from L. leichmannii.
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- Aberg A., Hahne S., Karlsson M., Larsson A., Ormö M., Ahgren A., Sjöberg B. M. Evidence for two different classes of redox-active cysteines in ribonucleotide reductase of Escherichia coli. J Biol Chem. 1989 Jul 25;264(21):12249–12252. [PubMed] [Google Scholar]
- Aebersold R. H., Leavitt J., Saavedra R. A., Hood L. E., Kent S. B. Internal amino acid sequence analysis of proteins separated by one- or two-dimensional gel electrophoresis after in situ protease digestion on nitrocellulose. Proc Natl Acad Sci U S A. 1987 Oct;84(20):6970–6974. doi: 10.1073/pnas.84.20.6970. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ashley G. W., Harris G., Stubbe J. The mechanism of Lactobacillus leichmannii ribonucleotide reductase. Evidence for 3' carbon-hydrogen bond cleavage and a unique role for coenzyme B12. J Biol Chem. 1986 Mar 25;261(9):3958–3964. [PubMed] [Google Scholar]
- BLAKLEY R. L. COBAMIDES AND RIBONUCLEOTIDE REDUCTION. I. COBAMIDE STIMULATION OF RIBONUCLEOTIDE REDUCTION IN EXTRACTS OF LACTOBACILLUS LEICHMANNII. J Biol Chem. 1965 May;240:2173–2180. [PubMed] [Google Scholar]
- Blakley R. L. Ribonucleoside triphosphate reductase from Lactobacillus leichmannii. Methods Enzymol. 1978;51:246–259. doi: 10.1016/s0076-6879(78)51034-3. [DOI] [PubMed] [Google Scholar]
- Chassy B. M., Giuffrida A. Method for the lysis of Gram-positive, asporogenous bacteria with lysozyme. Appl Environ Microbiol. 1980 Jan;39(1):153–158. doi: 10.1128/aem.39.1.153-158.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- EFTHYMIOU C., HANSEN P. A. An antigenic analysis of Lactobacillus acidophilus. J Infect Dis. 1962 May-Jun;110:258–267. doi: 10.1093/infdis/110.3.258. [DOI] [PubMed] [Google Scholar]
- Faust L. R., Connor J. A., Roof D. M., Hoch J. A., Babior B. M. Cloning, sequencing, and expression of the genes encoding the adenosylcobalamin-dependent ethanolamine ammonia-lyase of Salmonella typhimurium. J Biol Chem. 1990 Jul 25;265(21):12462–12466. [PubMed] [Google Scholar]
- Lane W. S., Galat A., Harding M. W., Schreiber S. L. Complete amino acid sequence of the FK506 and rapamycin binding protein, FKBP, isolated from calf thymus. J Protein Chem. 1991 Apr;10(2):151–160. doi: 10.1007/BF01024778. [DOI] [PubMed] [Google Scholar]
- Ledley F. D., Lumetta M., Nguyen P. N., Kolhouse J. F., Allen R. H. Molecular cloning of L-methylmalonyl-CoA mutase: gene transfer and analysis of mut cell lines. Proc Natl Acad Sci U S A. 1988 May;85(10):3518–3521. doi: 10.1073/pnas.85.10.3518. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lin A. N., Ashley G. W., Stubbe J. Location of the redox-active thiols of ribonucleotide reductase: sequence similarity between the Escherichia coli and Lactobacillus leichmannii enzymes. Biochemistry. 1987 Nov 3;26(22):6905–6909. doi: 10.1021/bi00396a006. [DOI] [PubMed] [Google Scholar]
- Lunn C. A., Kathju S., Wallace B. J., Kushner S. R., Pigiet V. Amplification and purification of plasmid-encoded thioredoxin from Escherichia coli K12. J Biol Chem. 1984 Aug 25;259(16):10469–10474. [PubMed] [Google Scholar]
- MacFerrin K. D., Terranova M. P., Schreiber S. L., Verdine G. L. Overproduction and dissection of proteins by the expression-cassette polymerase chain reaction. Proc Natl Acad Sci U S A. 1990 Mar;87(5):1937–1941. doi: 10.1073/pnas.87.5.1937. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mao S. S., Holler T. P., Bollinger J. M., Jr, Yu G. X., Johnston M. I., Stubbe J. Interaction of C225SR1 mutant subunit of ribonucleotide reductase with R2 and nucleoside diphosphates: tales of a suicidal enzyme. Biochemistry. 1992 Oct 13;31(40):9744–9751. doi: 10.1021/bi00155a030. [DOI] [PubMed] [Google Scholar]
- Mao S. S., Holler T. P., Yu G. X., Bollinger J. M., Jr, Booker S., Johnston M. I., Stubbe J. A model for the role of multiple cysteine residues involved in ribonucleotide reduction: amazing and still confusing. Biochemistry. 1992 Oct 13;31(40):9733–9743. doi: 10.1021/bi00155a029. [DOI] [PubMed] [Google Scholar]
- Mao S. S., Johnston M. I., Bollinger J. M., Stubbe J. Mechanism-based inhibition of a mutant Escherichia coli ribonucleotide reductase (cysteine-225----serine) by its substrate CDP. Proc Natl Acad Sci U S A. 1989 Mar;86(5):1485–1489. doi: 10.1073/pnas.86.5.1485. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mao S. S., Yu G. X., Chalfoun D., Stubbe J. Characterization of C439SR1, a mutant of Escherichia coli ribonucleotide diphosphate reductase: evidence that C439 is a residue essential for nucleotide reduction and C439SR1 is a protein possessing novel thioredoxin-like activity. Biochemistry. 1992 Oct 13;31(40):9752–9759. doi: 10.1021/bi00155a031. [DOI] [PubMed] [Google Scholar]
- Marsh E. N., McKie N., Davis N. K., Leadlay P. F. Cloning and structural characterization of the genes coding for adenosylcobalamin-dependent methylmalonyl-CoA mutase from Propionibacterium shermanii. Biochem J. 1989 Jun 1;260(2):345–352. doi: 10.1042/bj2600345. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Miura N., Ohtsuka E., Yamaberi N., Ikehara M., Uchida T., Okada Y. Use of the deoxyinosine-containing probe to isolate and sequence cDNA encoding the fusion (F) glycoprotein of Sendai virus (HVJ). Gene. 1985;38(1-3):271–274. doi: 10.1016/0378-1119(85)90228-8. [DOI] [PubMed] [Google Scholar]
- Nordlund P., Sjöberg B. M., Eklund H. Three-dimensional structure of the free radical protein of ribonucleotide reductase. Nature. 1990 Jun 14;345(6276):593–598. doi: 10.1038/345593a0. [DOI] [PubMed] [Google Scholar]
- Ohtsuka E., Matsuki S., Ikehara M., Takahashi Y., Matsubara K. An alternative approach to deoxyoligonucleotides as hybridization probes by insertion of deoxyinosine at ambiguous codon positions. J Biol Chem. 1985 Mar 10;260(5):2605–2608. [PubMed] [Google Scholar]
- Panagou D., Orr M. D., Dunstone J. R., Blakley R. L. A monomeric, allosteric enzyme with a single polypeptide chain. Ribonucleotide reductase of Lactobacillus leichmannii. Biochemistry. 1972 Jun 6;11(12):2378–2388. doi: 10.1021/bi00762a025. [DOI] [PubMed] [Google Scholar]
- Russel M., Model P. Direct cloning of the trxB gene that encodes thioredoxin reductase. J Bacteriol. 1985 Jul;163(1):238–242. doi: 10.1128/jb.163.1.238-242.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Singh D., Tamao Y., Blakley R. L. Allosterism, regulation and cooperativity: the case of ribonucleotide reductase of Lactobacillus leichmannii. Adv Enzyme Regul. 1976;15:81–100. doi: 10.1016/0065-2571(77)90010-3. [DOI] [PubMed] [Google Scholar]
- Stubbe J. Ribonucleotide reductases. Adv Enzymol Relat Areas Mol Biol. 1990;63:349–419. doi: 10.1002/9780470123096.ch6. [DOI] [PubMed] [Google Scholar]
- Stubbe J. Ribonucleotide reductases: amazing and confusing. J Biol Chem. 1990 Apr 5;265(10):5329–5332. [PubMed] [Google Scholar]
- Takahashi Y., Kato K., Hayashizaki Y., Wakabayashi T., Ohtsuka E., Matsuki S., Ikehara M., Matsubara K. Molecular cloning of the human cholecystokinin gene by use of a synthetic probe containing deoxyinosine. Proc Natl Acad Sci U S A. 1985 Apr;82(7):1931–1935. doi: 10.1073/pnas.82.7.1931. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thelander L., Reichard P. Reduction of ribonucleotides. Annu Rev Biochem. 1979;48:133–158. doi: 10.1146/annurev.bi.48.070179.001025. [DOI] [PubMed] [Google Scholar]