Skip to main content
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
. 1985 Mar;82(6):1658–1662. doi: 10.1073/pnas.82.6.1658

Stabilization of a primary loop in myosin subfragment 1 with a fluorescent crosslinker.

D Mornet, K Ue, M F Morales
PMCID: PMC397331  PMID: 3856845

Abstract

A bifunctional fluorescent alkylating agent, dibromobimane, has been used to stabilize a preexisting primary loop in myosin subfragment 1 (S-1). The crosslink achieved joins Cys-707 (called sulfhydryl group "SH1") of the 20-kDa domain (formerly called "20K" domain) with a thiol of the 50-kDa domain and seems to place the dibromobimane near the ATP-perturbable tryptophan.

Full text

PDF
1661

Images in this article

Selected References

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

  1. Applegate D., Reisler E. Protease-sensitive regions in myosin subfragment 1. Proc Natl Acad Sci U S A. 1983 Dec;80(23):7109–7112. doi: 10.1073/pnas.80.23.7109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bornstein P., Balian G. Cleavage at Asn-Gly bonds with hydroxylamine. Methods Enzymol. 1977;47:132–145. doi: 10.1016/0076-6879(77)47016-2. [DOI] [PubMed] [Google Scholar]
  3. Botts J., Takashi R., Torgerson P., Hozumi T., Muhlrad A., Mornet D., Morales M. F. On the mechanism of energy transduction in myosin subfragment 1. Proc Natl Acad Sci U S A. 1984 Apr;81(7):2060–2064. doi: 10.1073/pnas.81.7.2060. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Botts J., Ue K., Hozumi T., Samet J. Consequences of reacting the thiols of myosin subfragment 1. Biochemistry. 1979 Nov 13;18(23):5157–5163. doi: 10.1021/bi00590a020. [DOI] [PubMed] [Google Scholar]
  5. Bálint M., Wolf I., Tarcsafalvi A., Gergely J., Sréter F. A. Location of SH-1 and SH-2 in the heavy chain segment of heavy meromyosin. Arch Biochem Biophys. 1978 Oct;190(2):793–799. doi: 10.1016/0003-9861(78)90339-9. [DOI] [PubMed] [Google Scholar]
  6. Eisenberg E., Kielley W. W. Troponin-tropomyosin complex. Column chromatographic separation and activity of the three, active troponin components with and without tropomyosin present. J Biol Chem. 1974 Aug 10;249(15):4742–4748. [PubMed] [Google Scholar]
  7. Hiratsuka T. Direct cross-linking of three domains in the myosin head. J Biochem. 1984 Jul;96(1):269–272. doi: 10.1093/oxfordjournals.jbchem.a134823. [DOI] [PubMed] [Google Scholar]
  8. Hozumi T. Structure and function of myosin subfragment 1 as studied by tryptic digestion. Biochemistry. 1983 Feb 15;22(4):799–804. doi: 10.1021/bi00273a014. [DOI] [PubMed] [Google Scholar]
  9. Katoh T., Imae S., Morita F. Binding of F-actin to a region between SH1 and SH2 groups of myosin subfragment-1 which may determine the high affinity of acto-subfragment-1 complex at rigor. J Biochem. 1984 Feb;95(2):447–454. doi: 10.1093/oxfordjournals.jbchem.a134626. [DOI] [PubMed] [Google Scholar]
  10. Kosower N. S., Kosower E. M., Newton G. L., Ranney H. M. Bimane fluorescent labels: labeling of normal human red cells under physiological conditions. Proc Natl Acad Sci U S A. 1979 Jul;76(7):3382–3386. doi: 10.1073/pnas.76.7.3382. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Labbé J. P., Bertrand R., Audemard E., Kassab R., Walzthöny D., Wallimann T. The interaction of skeletal myosin subfragment 1 with the polyanion, heparin. Eur J Biochem. 1984 Sep 3;143(2):315–322. doi: 10.1111/j.1432-1033.1984.tb08374.x. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Lowey S., Slayter H. S., Weeds A. G., Baker H. Substructure of the myosin molecule. I. Subfragments of myosin by enzymic degradation. J Mol Biol. 1969 May 28;42(1):1–29. doi: 10.1016/0022-2836(69)90483-5. [DOI] [PubMed] [Google Scholar]
  14. Mornet D., Bertrand R., Pantel P., Audemard E., Kassab R. Structure of the actin-myosin interface. Nature. 1981 Jul 23;292(5821):301–306. doi: 10.1038/292301a0. [DOI] [PubMed] [Google Scholar]
  15. Mornet D., Pantel P., Audemard E., Kassab R. The limited tryptic cleavage of chymotryptic S-1: an approach to the characterization of the actin site in myosin heads. Biochem Biophys Res Commun. 1979 Aug 13;89(3):925–932. doi: 10.1016/0006-291x(79)91867-9. [DOI] [PubMed] [Google Scholar]
  16. Mornet D., Ue K., Morales M. F. Proteolysis and the domain organization of myosin subfragment 1. Proc Natl Acad Sci U S A. 1984 Feb;81(3):736–739. doi: 10.1073/pnas.81.3.736. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Mornet D., Ue K. Proteolysis and structure of skeletal muscle actin. Proc Natl Acad Sci U S A. 1984 Jun;81(12):3680–3684. doi: 10.1073/pnas.81.12.3680. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Offer G., Moos C., Starr R. A new protein of the thick filaments of vertebrate skeletal myofibrils. Extractions, purification and characterization. J Mol Biol. 1973 Mar 15;74(4):653–676. doi: 10.1016/0022-2836(73)90055-7. [DOI] [PubMed] [Google Scholar]
  19. Pope B. J., Wagner P. D., Weeds A. G. Heterogeneity of myosin heavy chains in subfragment-1 isoenzymes rabbit skeletal myosin. J Mol Biol. 1977 Jan 25;109(3):470–473. doi: 10.1016/s0022-2836(77)80024-7. [DOI] [PubMed] [Google Scholar]
  20. Reisler E. Sulfhydryl modification and labeling of myosin. Methods Enzymol. 1982;85(Pt B):84–93. doi: 10.1016/0076-6879(82)85012-x. [DOI] [PubMed] [Google Scholar]
  21. Rich S. A., Estes J. E. Detection of conformational changes in actin by proteolytic digestion: evidence for a new monomeric species. J Mol Biol. 1976 Jul 15;104(4):777–792. doi: 10.1016/0022-2836(76)90181-9. [DOI] [PubMed] [Google Scholar]
  22. Studier F. W. Analysis of bacteriophage T7 early RNAs and proteins on slab gels. J Mol Biol. 1973 Sep 15;79(2):237–248. doi: 10.1016/0022-2836(73)90003-x. [DOI] [PubMed] [Google Scholar]
  23. Sutoh K. Location of SH1 and SH2 along a heavy chain of myosin subfragment 1. Biochemistry. 1981 May 26;20(11):3281–3285. doi: 10.1021/bi00514a046. [DOI] [PubMed] [Google Scholar]
  24. Tong S. W., Elzinga M. The sequence of the NH2-terminal 204-residue fragment of the heavy chain of rabbit skeletal muscle myosin. J Biol Chem. 1983 Nov 10;258(21):13100–13110. [PubMed] [Google Scholar]
  25. Weeds A. G., Taylor R. S. Separation of subfragment-1 isoenzymes from rabbit skeletal muscle myosin. Nature. 1975 Sep 4;257(5521):54–56. doi: 10.1038/257054a0. [DOI] [PubMed] [Google Scholar]
  26. Wells J. A., Yount R. G. Active site trapping of nucleotides by crosslinking two sulfhydryls in myosin subfragment 1. Proc Natl Acad Sci U S A. 1979 Oct;76(10):4966–4970. doi: 10.1073/pnas.76.10.4966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Wells J. A., Yount R. G. Chemical modification of myosin by active-site trapping of metal-nucleotides with thiol crosslinking reagents. Methods Enzymol. 1982;85(Pt B):93–116. doi: 10.1016/0076-6879(82)85013-1. [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