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. 1995 Sep 1;130(5):1117–1125. doi: 10.1083/jcb.130.5.1117

Crystal structure and subunit dynamics of the abalone sperm lysin dimer: egg envelopes dissociate dimers, the monomer is the active species

PMCID: PMC2120550  PMID: 7657696

Abstract

Lysin is a 16-kD acrosomal protein used by abalone spermatozoa to create a hole in the egg vitelline envelope (VE) by a nonenzymatic mechanism. The crystal structure of the lysin monomer is known at 1.9 A resolution. The surface of the molecule reveals two tracks of basic residues running the length of one surface of the molecule and a patch of solvent-exposed hydrophobic residues on the opposite surface. Here we report that lysin dimerizes via interaction of the hydrophobic patches of monomers. Triton X-100 dissociates the dimer. The crystal structure of the dimer is described at 2.75 A resolution. Fluorescence energy transfer experiments show that the dimer has an approximate KD of 1 microM and that monomers exchange rapidly between dimers. Addition of isolated egg VE dissociates dimers, implicating monomers as the active species in the dissolution reaction. This work represents the first step in the elucidation of the mechanism by which lysin enables abalone spermatozoa to create a hole in the egg envelope during fertilization.

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Selected References

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  1. Adams S. R., Harootunian A. T., Buechler Y. J., Taylor S. S., Tsien R. Y. Fluorescence ratio imaging of cyclic AMP in single cells. Nature. 1991 Feb 21;349(6311):694–697. doi: 10.1038/349694a0. [DOI] [PubMed] [Google Scholar]
  2. Brünger A. T., Kuriyan J., Karplus M. Crystallographic R factor refinement by molecular dynamics. Science. 1987 Jan 23;235(4787):458–460. doi: 10.1126/science.235.4787.458. [DOI] [PubMed] [Google Scholar]
  3. Chacko S., Silverton E., Kam-Morgan L., Smith-Gill S., Cohen G., Davies D. Structure of an antibody-lysozyme complex unexpected effect of conservative mutation. J Mol Biol. 1995 Jan 20;245(3):261–274. doi: 10.1006/jmbi.1994.0022. [DOI] [PubMed] [Google Scholar]
  4. Clackson T., Wells J. A. A hot spot of binding energy in a hormone-receptor interface. Science. 1995 Jan 20;267(5196):383–386. doi: 10.1126/science.7529940. [DOI] [PubMed] [Google Scholar]
  5. Connolly M. L. Solvent-accessible surfaces of proteins and nucleic acids. Science. 1983 Aug 19;221(4612):709–713. doi: 10.1126/science.6879170. [DOI] [PubMed] [Google Scholar]
  6. Diller T. C., Shaw A., Stura E. A., Vacquier V. D., Stout C. D. Acid pH crystallization of the basic protein lysin from the spermatozoa of red abalone (Haliotis rufescens). Acta Crystallogr D Biol Crystallogr. 1994 Jul 1;50(Pt 4):620–626. doi: 10.1107/S0907444993013356. [DOI] [PubMed] [Google Scholar]
  7. Hong K., Vacquier V. D. Fusion of liposomes induced by a cationic protein from the acrosome granule of abalone spermatozoa. Biochemistry. 1986 Feb 11;25(3):543–549. doi: 10.1021/bi00351a004. [DOI] [PubMed] [Google Scholar]
  8. Janin J., Chothia C. The structure of protein-protein recognition sites. J Biol Chem. 1990 Sep 25;265(27):16027–16030. [PubMed] [Google Scholar]
  9. 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]
  10. Lee Y. H., Ota T., Vacquier V. D. Positive selection is a general phenomenon in the evolution of abalone sperm lysin. Mol Biol Evol. 1995 Mar;12(2):231–238. doi: 10.1093/oxfordjournals.molbev.a040200. [DOI] [PubMed] [Google Scholar]
  11. Lewis C. A., Leighton D. L., Vacquier V. D. Morphology of abalone spermatozoa before and after the acrosome reaction. J Ultrastruct Res. 1980 Jul;72(1):39–46. doi: 10.1016/s0022-5320(80)90133-1. [DOI] [PubMed] [Google Scholar]
  12. Lewis C. A., Talbot C. F., Vacquier V. D. A protein from abalone sperm dissolves the egg vitelline layer by a nonenzymatic mechanism. Dev Biol. 1982 Jul;92(1):227–239. doi: 10.1016/0012-1606(82)90167-1. [DOI] [PubMed] [Google Scholar]
  13. Ren Z., Meyer T., McRee D. E. Atomic structure of a cytochrome c' with an unusual ligand-controlled dimer dissociation at 1.8 A resolution. J Mol Biol. 1993 Nov 20;234(2):433–445. doi: 10.1006/jmbi.1993.1597. [DOI] [PubMed] [Google Scholar]
  14. Shaw A., McRee D. E., Vacquier V. D., Stout C. D. The crystal structure of lysin, a fertilization protein. Science. 1993 Dec 17;262(5141):1864–1867. doi: 10.1126/science.8266073. [DOI] [PubMed] [Google Scholar]
  15. Vacquier V. D., Carner K. R., Stout C. D. Species-specific sequences of abalone lysin, the sperm protein that creates a hole in the egg envelope. Proc Natl Acad Sci U S A. 1990 Aug;87(15):5792–5796. doi: 10.1073/pnas.87.15.5792. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Vacquier V. D., Lee Y. H. Abalone sperm lysin: unusual mode of evolution of a gamete recognition protein. Zygote. 1993 Aug;1(3):181–196. doi: 10.1017/s0967199400001465. [DOI] [PubMed] [Google Scholar]

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