Abstract

The purpose of the research is to gain an understanding of the mechanism of self-assembly of tubulin and the control of microtubule formation and depolymerization. In these studies we are examining the polymorphism of tubulin self-assembly by determining the effects of a number of variables on the protofilament number of formed microtubules and on the formation of complex ribbons and sheets of protofilaments. We are also investigating the types of intermediates which are present during self-assembly of 6S tubulin. The roles of sulfhydryl and disulfide bonds in tubulin are being examined. These will be modified specifically with agents such as fluorodinitrobenzene, N-ethylmaleimide, dansyl chloride, dansylaziridine, and 2-mercaptoethanol. Effects on the physical structure of tubulin by these agents will be determined by difference spectroscopy, fluorimetric and circular dichroism spectroscopy studies. The function of the tightly bound metal ion is also being examined. The effects of exchanging bound Mg with Mn, Zn, and Co on the physical structure of tubulin and the self-assembly reaction will be investigated.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM036953-11
Application #
3291675
Study Section
Biophysics and Biophysical Chemistry A Study Section (BBCA)
Project Start
1986-09-01
Project End
1991-08-31
Budget Start
1987-09-01
Budget End
1988-08-31
Support Year
11
Fiscal Year
1987
Total Cost
Indirect Cost

  Institution

Name
University of Kansas Lawrence
Department
Type
Schools of Arts and Sciences
DUNS #
072933393
City
Lawrence
State
KS
Country
United States
Zip Code
66045

  Publications

Singer, W D; Parker, S K; Himes, R H et al. (1994) Polymerization of Antarctic fish tubulins at low temperatures: role of carboxy-terminal domains. Biochemistry 33:15389-96
Shivanna, B D; Mejillano, M R; Williams, T D et al. (1993) Exchangeable GTP binding site of beta-tubulin. Identification of cysteine 12 as the major site of cross-linking by direct photoaffinity labeling. J Biol Chem 268:127-32
Singer, W D; Himes, R H (1992) Cellular uptake and tubulin binding properties of four Vinca alkaloids. Biochem Pharmacol 43:545-51
Reed, J; Hull, W E; Ponstingl, H et al. (1992) Conformational properties of the beta(400-436) and beta(400-445) C-terminal peptides of porcine brain tubulin. Biochemistry 31:11888-95
Mathew, A E; Mejillano, M R; Nath, J P et al. (1992) Synthesis and evaluation of some water-soluble prodrugs and derivatives of taxol with antitumor activity. J Med Chem 35:145-51
Mejillano, M R; Tolo, E T; Williams Jr, R C et al. (1992) The conversion of tubulin carboxyl groups to amides has a stabilizing effect on microtubules. Biochemistry 31:3478-83
Mejillano, M R; Himes, R H (1991) Binding of guanine nucleotides and Mg2+ to tubulin with a nucleotide-depleted exchangeable site. Arch Biochem Biophys 291:356-62
Osei, A A; Everett, G W; Ringel, I et al. (1991) The interaction of [13C]-enriched colchicine with tubulin as determined by NMR spectroscopy. Biochim Biophys Acta 1078:339-44
Mejillano, M R; Himes, R H (1991) Assembly properties of tubulin after carboxyl group modification. J Biol Chem 266:657-64
Osei, A A; Everett, G W; Himes, R H (1990) Evidence that the tightly bound magnesium in tubulin is associated with the N-site GTP. FEBS Lett 276:85-7

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