Multiple sites for subtilisin cleavage of tubulin: Effects of divalent cations

Lobert, Sharon; Hennington, Bettye Sue; Correia, John J.

doi:10.1002/cm.970250308

Cited by 26 publications

(16 citation statements)

References 53 publications

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“…29,45 We looked at both the reaction intermediates as well as the products generated by depolymerization of GMPCPP-stabilized SMTs to determine whether there was a difference in the reaction intermediates between SMTs and normal MTs as intermediate formation or oligomeric reaction products could be enhanced by subtilisin treatment. 46 In the presence of MgAMPPNP, MCAK induced robust protofilament peels on many of the GMPCPP-stabilized MT ends ( Fig. 3A and B).…”

Section: Resultsmentioning

confidence: 93%

The C-termini of tubulin and the specific geometry of tubulin substrates influence the depolymerization activity of MCAK

Hertzer

Walczak²

2008

Cell Cycle

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Section: Resultsmentioning

confidence: 93%

The C-termini of tubulin and the specific geometry of tubulin substrates influence the depolymerization activity of MCAK

Hertzer

Walczak²

2008

Cell Cycle

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“…The interaction of Ca 2ϩ with the Cterminal part of tubulin was suggested by the fact that the direct inhibition of tubulin assembly by Ca 2ϩ disappears when using subtilisin-treated tubulin. A direct binding of Ca 2ϩ on proteolytic fragments of the ␣-or ␤-tubulin tails was also demonstrated (45,58), and a high affinity Ca 2ϩ binding site was finally proposed in the 423-446 region of the tubulin tail with a K d evaluated to be 11 M (59).…”

Section: Discussionmentioning

confidence: 99%

The C Terminus of Tubulin, a Versatile Partner for Cationic Molecules

Lefèvre¹,

Chernov

Joshi

et al. 2011

Journal of Biological Chemistry

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The C-terminal region of tubulin is involved in multiple aspects of the regulation of microtubule assembly. To elucidate the molecular mechanisms of this regulation, we study here, using different approaches, the interaction of Tau Microtubules are involved in a number of critical cellular processes, such as the determination of cell shape, chromosome segregation, intracellular transport of vesicles and organelles, and cell migration. Microtubules consist mainly of ␣␤-tubulin heterodimers organized head-to-tail into protofilaments whose parallel self-association gives rise to microtubules (1-4). ␣-and ␤-tubulin monomers have each a molecular mass of 50 kDa and are organized in three domains, namely the N-terminal domain (amino acid residues 1-205) involved in nucleotide binding, the intermediate domain (amino acid residues 206 -384), and the C-terminal domain (amino acid residue 385 to the C terminus) (5). The C-terminal domain of tubulin represents a critical part of the binding site of different tubulin/microtubules partners, such as MAPs, 4 which are major regulators of microtubule dynamics (6 -8), or polycations, which promote tubulin assembly in vitro in different polymeric forms (9). The C-terminal domain comprises a highly negatively charged tail of about 20 amino acid residues (named herein the C-terminal tail (CTT)), which protrudes from the surface of microtubules. In agreement with its participation in the regulation of microtubule assembly through interactions with partners, the CTT is also the most divergent part of tubulin, and variations among tubulin isotypes (10) may explain the modulation of the dynamics of microtubule assembly in specific tissues or cytoplasmic regions.Different structure information has been obtained regarding the C-terminal domain of tubulin by using either fulllength tubulin or peptide fragments. Electron crystallography of zinc-induced tubulin sheets showed the presence of two anti-parallel ␣-helices (helix H11 (amino acid residues 385-397) and helix H12 (amino acid residues 418 -433)) lying at the outer surface of tubulin. The regions corresponding to the CTTs of either ␣-or ␤-tubulin were however not observed, probably due to the flexibility of this part of the protein (5). These observations were confirmed by x-ray diffraction analyses of crystal complexes formed between tubulin and the RB3-stathmin-like domain (11-13). Other structural data were obtained with peptides from the C-terminal region of tubulin studied either when free in solution or in interaction with different partners. NMR structure investigations on ␣-and ␤-tubulin C-terminal peptides showed that both ␣ (residues 404 -451) and ␤ (residues 394 -445) peptides have no defined secondary structure in aqueous solution but contain a well □ S The on-line version of this article (available at http://www.jbc.org) contains supplemental 4 The abbreviations used are: MAP, microtubule-associated protein; ␣Tub410C, amino acid residues 410 -451 from ␣1a-tubulin; CTT, tubulin C-terminal tail; ITC, isothermal titration ...

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“…The structure contains information at 3.7 Å on all but the final 10-18 carboxyl tail residues [35,36]. The disordered carboxyl tail is the site for subtilisin digestion [37], numerous post-translational modifications [38,39] and MAP binding [40,41]. Since zinc sheets are composed of protofilaments, the structure contains information on the polymerized longitudinal contacts both within and between dimers.…”

Section: Implications Of the Structure Of Tubulin For Drug Binding Anmentioning

confidence: 99%

Physiochemical Aspects of Tubulin-Interacting Antimitotic Drugs

Lobert²

2001

CPD

102

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A diverse group of natural biological compounds bind to microtubules and suppress microtubule dynamics. Here we review the mechanism of microtubule assembly and dynamics as well as structural features that are important for nucleotide binding, GTP hydrolysis and stabilization of longitudinal and lateral protofilament contacts. Specific emphasis is placed upon the polar structure of the microtubule, the exposure of the nucleotide hydrolysis site at the + end and the conformational and configurational plasticity of the microtubule lattice. These features have important implications for the mechanism of dynamic instability and the disruptive action of antimitotic drugs. We then discuss the various classes of tubulin binding drugs emphasizing their site and mode of binding as well as the structural and energetic basis for their effects on microtubule assembly and dynamics. A common feature of tubulin-interacting compounds is a linkage to assembly, either the stabilization of a microtubule lattice by compounds like taxol or epothilone A, or the preferential formation of alternate lattice contacts and polymers at microtubule ends by compounds like colchicine, vinca alkaloids and cryptophycin-52. Finally, we explore the likely possibility that these drugs also disrupt the regulation of microtubule dynamics. Future generations of these compounds may be selectively developed to directly target the proteins that regulate mitotic spindle dynamics.

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Multiple sites for subtilisin cleavage of tubulin: Effects of divalent cations

Cited by 26 publications

References 53 publications

The C-termini of tubulin and the specific geometry of tubulin substrates influence the depolymerization activity of MCAK

The C-termini of tubulin and the specific geometry of tubulin substrates influence the depolymerization activity of MCAK

The C Terminus of Tubulin, a Versatile Partner for Cationic Molecules

Physiochemical Aspects of Tubulin-Interacting Antimitotic Drugs

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