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 Mar 15;89(6):2071–2075. doi: 10.1073/pnas.89.6.2071

Primary structure of Euplotes raikovi pheromones: comparison of five sequences of pheromones from cells with variable mating interactions.

S Raffioni 1, C Miceli 1, A Vallesi 1, S K Chowdhury 1, B T Chait 1, P Luporini 1, R A Bradshaw 1
PMCID: PMC48598  PMID: 1549567

Abstract

The amino acid sequences of five pheromones, Er-2, Er-3, Er-9, Er-11, and Er-20, secreted by cells of different mating types of the ciliated protozoa Euplotes raikovi, have been determined by automated Edman analyses of the whole proteins and germane fragments. In each case, the molecular mass was determined by plasma desorption or laser desorption mass spectrometry and was in excellent agreement with the calculated values. Where available, the determined sequences were also in accord with the corresponding segments of the precursor molecules predicted from relevant nucleic acid sequences. Of the five, two were found to be identical (Er-2 and Er-9) and one (Er-3) was identical to a pheromone previously sequenced (Er-1), even though mating pair formation was found to take place (although to a limited extent) when cells secreting those pheromones were combined in a mixture. Comparison of the five unique sequences suggested a closer relationship between Er-1 (Er-3) and Er-10 and between Er-11 and Er-20 (44% and 56% identity, respectively) than was generally observed among the other members. This pairing was also supported by hydrophobicity analyses. Interestingly, Er-20 cannot, as a rule, induce cell union in any of the other cell types, including cells secreting Er-11, despite the fact that Er-20 and Er-11 are the most similar of the five unique sequences. Thus sequence identity and secondary structure profiles are not a good indicator of biological relatedness as manifested in heterologous receptor interaction.

Full text

PDF
2073

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. Factors affecting the ultraviolet laser desorption of proteins. Rapid Commun Mass Spectrom. 1989 Jul;3(7):233–237. doi: 10.1002/rcm.1290030708. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. 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]
  5. Hayashi R., Moore S., Merrifield R. B. Preparation of pancreatic ribonucleases 1-114 and 1-115 and their reactivation by mixture and synthetic COOH-terminal peptides. J Biol Chem. 1973 Jun 10;248(11):3889–3892. [PubMed] [Google Scholar]
  6. Heinrikson R. L., Meredith S. C. Amino acid analysis by reverse-phase high-performance liquid chromatography: precolumn derivatization with phenylisothiocyanate. Anal Biochem. 1984 Jan;136(1):65–74. doi: 10.1016/0003-2697(84)90307-5. [DOI] [PubMed] [Google Scholar]
  7. Hopp T. P., Woods K. R. Prediction of protein antigenic determinants from amino acid sequences. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3824–3828. doi: 10.1073/pnas.78.6.3824. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. James R., Bradshaw R. A. Polypeptide growth factors. Annu Rev Biochem. 1984;53:259–292. doi: 10.1146/annurev.bi.53.070184.001355. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Miceli C., Concetti A., Luporini P. Isolation of the mating-inducing factor of the ciliate Euplotes. Exp Cell Res. 1983 Dec;149(2):593–598. doi: 10.1016/0014-4827(83)90371-3. [DOI] [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. Miceli C., La Terza A., Melli M. Isolation and structural characterization of cDNA clones encoding the mating pheromone Er-1 secreted by the ciliate Euplotes raikovi. Proc Natl Acad Sci U S A. 1989 May;86(9):3016–3020. doi: 10.1073/pnas.86.9.3016. [DOI] [PMC free article] [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. 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]
  16. 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]
  17. 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]
  18. 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 Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES