Skip to main content
PMC home page
Protein Science : A Publication of the Protein Society logoLink to Protein Science : A Publication of the Protein Society
. 1992 Jun;1(6):777–785. doi: 10.1002/pro.5560010609

The disulfide bond pairing of the pheromones Er-1 and Er-2 of the ciliated protozoan Euplotes raikovi.

A E Stewart 1, S Raffioni 1, T Chaudhary 1, B T Chait 1, P Luporini 1, R A Bradshaw 1
PMCID: PMC2142243  PMID: 1304918

Abstract

The disulfide pairings of the two Euplotes raikovi pheromones Er-1 and Er-2 have been determined by chemical and mass spectrometric analyses. Cystine-linked peptides from thermolytic digestions of the native molecules were purified by reverse-phase high performance liquid chromatography and identified in the known sequences to make the assignments. The same pairing, Cys(I)-Cys(IV), Cys(II)-Cys(VI), and Cys(III)-Cys(V), was found in both pheromones, suggesting that this pattern occurs commonly throughout this family of molecules. This arrangement of disulfides indicates that the three-dimensional structure is defined by three loops, which can vary in size and charge distribution from one pheromone to another.

Full Text

The Full Text of this article is available as a PDF (739.7 KB).

Selected References

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

  1. Anderson D., Raffioni S., Luporini P., Bradshaw R. A., Eisenberg D. Crystallization of the Euplotes raikovi mating pheromone Er-1. J Mol Biol. 1990 Nov 5;216(1):1–2. doi: 10.1016/S0022-2836(05)80055-5. [DOI] [PubMed] [Google Scholar]
  2. Beavis R. C., Chait B. T. High-accuracy molecular mass determination of proteins using matrix-assisted laser desorption mass spectrometry. Anal Chem. 1990 Sep 1;62(17):1836–1840. doi: 10.1021/ac00216a020. [DOI] [PubMed] [Google Scholar]
  3. Bradshaw R. A., Babin D. R., Nomoto M., Srinivasin N. G., Ericsson L. H., Walsh K. A., Neurath H. The amino acid sequence of bovine carboxypeptidase A. II. Tryptic and chymotryptic peptides of the cyanogen bromide fragment F-III. Biochemistry. 1969 Sep;8(9):3859–3871. doi: 10.1021/bi00837a052. [DOI] [PubMed] [Google Scholar]
  4. Chowdhury S. K., Katta V., Chait B. T. An electrospray-ionization mass spectrometer with new features. Rapid Commun Mass Spectrom. 1990 Mar;4(3):81–87. doi: 10.1002/rcm.1290040305. [DOI] [PubMed] [Google Scholar]
  5. Concetti A., Raffioni S., Miceli C., Barra D., Luporini P. Purification to apparent homogeneity of the mating pheromone of mat-1 homozygous Euplotes raikovi. J Biol Chem. 1986 Aug 15;261(23):10582–10586. [PubMed] [Google Scholar]
  6. Guy O., Shapanka R., Greene L. J. The structure of the bovine pancreatic secretory trypsin inhibitor--Kazal's inhibitor. 3. Determination of the disulfide bonds and proteolysis by thermolysin. J Biol Chem. 1971 Dec 25;246(24):7740–7747. [PubMed] [Google Scholar]
  7. Kubota T., Tokoroyama T., Tsukuda Y., Koyama H., Miyake A. Isolation and structure determination of blepharismin, a conjugation initiating gamone in the ciliate blepharisma. Science. 1973 Jan 26;179(4071):400–402. doi: 10.1126/science.179.4071.400. [DOI] [PubMed] [Google Scholar]
  8. Luporini P., Raffioni S., Concetti A., Miceli C. The ciliate Euplotes raikovi heterozygous at the mat genetic locus coreleases two individual species of mating pheromone: Genetic and biochemical evidence. Proc Natl Acad Sci U S A. 1986 May;83(9):2889–2893. doi: 10.1073/pnas.83.9.2889. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Matsubara H., Singer A., Sasaki R., Jukes T. H. Observations on the specificity of a thermostable bacterial protease "thermolysin". Biochem Biophys Res Commun. 1965 Nov 8;21(3):242–247. doi: 10.1016/0006-291x(65)90278-0. [DOI] [PubMed] [Google Scholar]
  10. Meyer F., Schmidt H. J., Plümper E., Hasilik A., Mersmann G., Meyer H. E., Engström A., Heckmann K. UGA is translated as cysteine in pheromone 3 of Euplotes octocarinatus. Proc Natl Acad Sci U S A. 1991 May 1;88(9):3758–3761. doi: 10.1073/pnas.88.9.3758. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Miceli C., La Terza A., Bradshaw R. A., Luporini P. Identification and structural characterization of a cDNA clone encoding a membrane-bound form of the polypeptide pheromone Er-1 in the ciliate protozoan Euplotes raikovi. Proc Natl Acad Sci U S A. 1992 Mar 1;89(5):1988–1992. doi: 10.1073/pnas.89.5.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Miceli C., La Terza A., Bradshaw R. A., Luporini P. Structural characterization of mating pheromone precursors of the ciliate protozoan Euplotes raikovi. High conservation of pre and pro regions versus high variability of secreted regions. Eur J Biochem. 1991 Dec 18;202(3):759–764. doi: 10.1111/j.1432-1033.1991.tb16430.x. [DOI] [PubMed] [Google Scholar]
  13. Miyake A., Beyer J. Blepharmone: a conjugation-inducing glycoprotein in the ciliate blepharisma. Science. 1974 Aug 16;185(4151):621–623. doi: 10.1126/science.185.4151.621. [DOI] [PubMed] [Google Scholar]
  14. Ortenzi C., Miceli C., Bradshaw R. A., Luporini P. Identification and initial characterization of an autocrine pheromone receptor in the protozoan ciliate Euplotes raikovi. J Cell Biol. 1990 Aug;111(2):607–614. doi: 10.1083/jcb.111.2.607. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Piszkiewicz D., Landon M., Smith E. L. Anomalous cleavage of aspartyl-proline peptide bonds during amino acid sequence determinations. Biochem Biophys Res Commun. 1970 Sep 10;40(5):1173–1178. doi: 10.1016/0006-291x(70)90918-6. [DOI] [PubMed] [Google Scholar]
  16. Raffioni S., Luporini P., Bradshaw R. A. Purification, characterization, and amino acid sequence of the mating pheromone Er-10 of the ciliate Euplotes raikovi. Biochemistry. 1989 Jun 13;28(12):5250–5256. doi: 10.1021/bi00438a049. [DOI] [PubMed] [Google Scholar]
  17. Raffioni S., Luporini P., Chait B. T., Disper S. S., Bradshaw R. A. Primary structure of the mating pheromone Er-1 of the ciliate Euplotes raikovi. J Biol Chem. 1988 Dec 5;263(34):18152–18159. [PubMed] [Google Scholar]
  18. Raffioni S., Miceli C., Concetti A., Barra D., Luporini P. Purification and characterization of new mating pheromones of the ciliate Euplotes raikovi. Exp Cell Res. 1987 Oct;172(2):417–424. doi: 10.1016/0014-4827(87)90399-5. [DOI] [PubMed] [Google Scholar]
  19. SPACKMAN D. H., STEIN W. H., MOORE S. The disulfide bonds of ribonuclease. J Biol Chem. 1960 Mar;235:648–659. [PubMed] [Google Scholar]
  20. Schulze Dieckhoff H., Freiburg M., Heckmann K. The isolation of gamones 3 and 4 of Euplotes octocarinatus. Eur J Biochem. 1987 Oct 1;168(1):89–94. doi: 10.1111/j.1432-1033.1987.tb13391.x. [DOI] [PubMed] [Google Scholar]

Articles from Protein Science : A Publication of the Protein Society are provided here courtesy of The Protein Society

RESOURCES