Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1992 Feb 15;89(4):1164–1168. doi: 10.1073/pnas.89.4.1164

Neisseria meningitidis encodes an FK506-inhibitable rotamase.

B A Sampson 1, E C Gotschlich 1
PMCID: PMC48409  PMID: 1371354

Abstract

Eukaryotic peptidyl-prolyl cis-trans isomerases (rotamases) fall into two classes, the cyclophilins inhibited by cyclosporin A and the FK506-binding proteins inhibited by the macrolide antibiotic FK506. In prokaryotes homologs of cyclophilins have been identified and found to have rotamase activity. Sequence similarities have been noted between FK506-binding proteins and gene products in a number of bacterial species, but whether these bacterial proteins have rotamase activity is not known. Using the polymerase chain reaction, we have cloned and sequenced a homolog of an FK506-binding protein from Neisseria meningitidis and expressed the gene product as a fusion protein with maltose-binding protein. The fusion protein was purified by affinity chromatography. By measuring the rate of chymotrypsin cleavage of the substrate succinyl-Ala-Ala-Pro-Phe p-nitroanilide, we found that the fusion protein had rotamase activity comparable to that of human FK506-binding protein. This rotamase activity was inhibited by FK506.

Full text

PDF
1166

Images in this article

1166Image
on p.1166

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aho E. L., Cannon J. G. Characterization of a silent pilin gene locus from Neisseria meningitidis strain FAM18. Microb Pathog. 1988 Nov;5(5):391–398. doi: 10.1016/0882-4010(88)90039-3. [DOI] [PubMed] [Google Scholar]
  2. Cianciotto N. P., Eisenstein B. I., Mody C. H., Engleberg N. C. A mutation in the mip gene results in an attenuation of Legionella pneumophila virulence. J Infect Dis. 1990 Jul;162(1):121–126. doi: 10.1093/infdis/162.1.121. [DOI] [PubMed] [Google Scholar]
  3. Cianciotto N. P., Eisenstein B. I., Mody C. H., Toews G. B., Engleberg N. C. A Legionella pneumophila gene encoding a species-specific surface protein potentiates initiation of intracellular infection. Infect Immun. 1989 Apr;57(4):1255–1262. doi: 10.1128/iai.57.4.1255-1262.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Engleberg N. C., Carter C., Weber D. R., Cianciotto N. P., Eisenstein B. I. DNA sequence of mip, a Legionella pneumophila gene associated with macrophage infectivity. Infect Immun. 1989 Apr;57(4):1263–1270. doi: 10.1128/iai.57.4.1263-1270.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fischer G., Wittmann-Liebold B., Lang K., Kiefhaber T., Schmid F. X. Cyclophilin and peptidyl-prolyl cis-trans isomerase are probably identical proteins. Nature. 1989 Feb 2;337(6206):476–478. doi: 10.1038/337476a0. [DOI] [PubMed] [Google Scholar]
  6. Flanagan W. M., Corthésy B., Bram R. J., Crabtree G. R. Nuclear association of a T-cell transcription factor blocked by FK-506 and cyclosporin A. Nature. 1991 Aug 29;352(6338):803–807. doi: 10.1038/352803a0. [DOI] [PubMed] [Google Scholar]
  7. Gill S. C., von Hippel P. H. Calculation of protein extinction coefficients from amino acid sequence data. Anal Biochem. 1989 Nov 1;182(2):319–326. doi: 10.1016/0003-2697(89)90602-7. [DOI] [PubMed] [Google Scholar]
  8. Harding M. W., Galat A., Uehling D. E., Schreiber S. L. A receptor for the immunosuppressant FK506 is a cis-trans peptidyl-prolyl isomerase. Nature. 1989 Oct 26;341(6244):758–760. doi: 10.1038/341758a0. [DOI] [PubMed] [Google Scholar]
  9. Harrison R. K., Stein R. L. Substrate specificities of the peptidyl prolyl cis-trans isomerase activities of cyclophilin and FK-506 binding protein: evidence for the existence of a family of distinct enzymes. Biochemistry. 1990 Apr 24;29(16):3813–3816. doi: 10.1021/bi00468a001. [DOI] [PubMed] [Google Scholar]
  10. Hayano T., Takahashi N., Kato S., Maki N., Suzuki M. Two distinct forms of peptidylprolyl-cis-trans-isomerase are expressed separately in periplasmic and cytoplasmic compartments of Escherichia coli cells. Biochemistry. 1991 Mar 26;30(12):3041–3048. doi: 10.1021/bi00226a009. [DOI] [PubMed] [Google Scholar]
  11. Kato J., Chu L., Kitano K., DeVault J. D., Kimbara K., Chakrabarty A. M., Misra T. K. Nucleotide sequence of a regulatory region controlling alginate synthesis in Pseudomonas aeruginosa: characterization of the algR2 gene. Gene. 1989 Dec 7;84(1):31–38. doi: 10.1016/0378-1119(89)90136-4. [DOI] [PubMed] [Google Scholar]
  12. Kawamukai M., Matsuda H., Fujii W., Utsumi R., Komano T. Nucleotide sequences of fic and fic-1 genes involved in cell filamentation induced by cyclic AMP in Escherichia coli. J Bacteriol. 1989 Aug;171(8):4525–4529. doi: 10.1128/jb.171.8.4525-4529.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  14. Liu J., Farmer J. D., Jr, Lane W. S., Friedman J., Weissman I., Schreiber S. L. Calcineurin is a common target of cyclophilin-cyclosporin A and FKBP-FK506 complexes. Cell. 1991 Aug 23;66(4):807–815. doi: 10.1016/0092-8674(91)90124-h. [DOI] [PubMed] [Google Scholar]
  15. Liu J., Walsh C. T. Peptidyl-prolyl cis-trans-isomerase from Escherichia coli: a periplasmic homolog of cyclophilin that is not inhibited by cyclosporin A. Proc Natl Acad Sci U S A. 1990 Jun;87(11):4028–4032. doi: 10.1073/pnas.87.11.4028. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lundemose A. G., Birkelund S., Fey S. J., Larsen P. M., Christiansen G. Chlamydia trachomatis contains a protein similar to the Legionella pneumophila mip gene product. Mol Microbiol. 1991 Jan;5(1):109–115. doi: 10.1111/j.1365-2958.1991.tb01831.x. [DOI] [PubMed] [Google Scholar]
  17. Michnick S. W., Rosen M. K., Wandless T. J., Karplus M., Schreiber S. L. Solution structure of FKBP, a rotamase enzyme and receptor for FK506 and rapamycin. Science. 1991 May 10;252(5007):836–839. doi: 10.1126/science.1709301. [DOI] [PubMed] [Google Scholar]
  18. Moore J. M., Peattie D. A., Fitzgibbon M. J., Thomson J. A. Solution structure of the major binding protein for the immunosuppressant FK506. Nature. 1991 May 16;351(6323):248–250. doi: 10.1038/351248a0. [DOI] [PubMed] [Google Scholar]
  19. Perry A. C., Nicolson I. J., Saunders J. R. Neisseria meningitidis C114 contains silent, truncated pilin genes that are homologous to Neisseria gonorrhoeae pil sequences. J Bacteriol. 1988 Apr;170(4):1691–1697. doi: 10.1128/jb.170.4.1691-1697.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Sawada S., Suzuki G., Kawase Y., Takaku F. Novel immunosuppressive agent, FK506. In vitro effects on the cloned T cell activation. J Immunol. 1987 Sep 15;139(6):1797–1803. [PubMed] [Google Scholar]
  23. Schreiber S. L. Chemistry and biology of the immunophilins and their immunosuppressive ligands. Science. 1991 Jan 18;251(4991):283–287. doi: 10.1126/science.1702904. [DOI] [PubMed] [Google Scholar]
  24. Siekierka J. J., Hung S. H., Poe M., Lin C. S., Sigal N. H. A cytosolic binding protein for the immunosuppressant FK506 has peptidyl-prolyl isomerase activity but is distinct from cyclophilin. Nature. 1989 Oct 26;341(6244):755–757. doi: 10.1038/341755a0. [DOI] [PubMed] [Google Scholar]
  25. Standaert R. F., Galat A., Verdine G. L., Schreiber S. L. Molecular cloning and overexpression of the human FK506-binding protein FKBP. Nature. 1990 Aug 16;346(6285):671–674. doi: 10.1038/346671a0. [DOI] [PubMed] [Google Scholar]
  26. Takahashi N., Hayano T., Suzuki M. Peptidyl-prolyl cis-trans isomerase is the cyclosporin A-binding protein cyclophilin. Nature. 1989 Feb 2;337(6206):473–475. doi: 10.1038/337473a0. [DOI] [PubMed] [Google Scholar]
  27. Tropschug M., Wachter E., Mayer S., Schönbrunner E. R., Schmid F. X. Isolation and sequence of an FK506-binding protein from N. crassa which catalyses protein folding. Nature. 1990 Aug 16;346(6285):674–677. doi: 10.1038/346674a0. [DOI] [PubMed] [Google Scholar]
  28. Van Duyne G. D., Standaert R. F., Karplus P. A., Schreiber S. L., Clardy J. Atomic structure of FKBP-FK506, an immunophilin-immunosuppressant complex. Science. 1991 May 10;252(5007):839–842. doi: 10.1126/science.1709302. [DOI] [PubMed] [Google Scholar]
  29. Wiederrecht G., Brizuela L., Elliston K., Sigal N. H., Siekierka J. J. FKB1 encodes a nonessential FK 506-binding protein in Saccharomyces cerevisiae and contains regions suggesting homology to the cyclophilins. Proc Natl Acad Sci U S A. 1991 Feb 1;88(3):1029–1033. doi: 10.1073/pnas.88.3.1029. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES