Differential effects of active isomers, segments, and analogs of dolastatin 10 on ligand interactions with tubulin. Correlation with cytotoxicity

Ruoli Bai, Mary Carmen Roach, Srirangam K. Jayaram, Jozef Barkoczy, George R. Pettit, Richard F. Luduena, Ernest Hamel

Research output: Contribution to journalArticle

76 Citations (Scopus)

Abstract

Dolastatin 10 is a potent antimitotic peptide isolated from the marine mollusk Dolabella auricularia. Four of its five residues are modified amino acids (in sequence, dolavaline, valine, dolaisoleuine, dolaproine, dolaphenine). Besides inhibiting tubulin polymerization, dolastatin 10 noncompetitively inhibits vinca alkaloid binding to tubulin, inhibits nucleotide exchange and formation of the βs cross-link, and stabilizes the colchicine binding activity of tubulin. To examine the mechanism of action of dolastatin 10 we prepared six chiral isomers, one tri- and one tetrapeptide segment, and one pentapeptide analog of dolastatin 10, all of which differ little from dolastatin 10 as inhibitors of tubulin polymerization. However, only two of the chiral isomers were similar to dolastatin 10 in their cytotoxicity for L1210 murine leukemia cells and in their effects on vinblastine binding, nucleotide exchange, βs cross-link formation, and colchicine binding. These were isomer 2, with reversal of configuration at position C(19a) in the dolaisoleuine moiety, and isomer 19, with reversal of configuration at position C(6) in the dolaphenine moiety. The pentapeptides with reduced cytotoxicity and reduced effects on tubulin interactions with other ligands were all modified in the dolaproine moiety at positions C(9) and/ or C(10). The tripeptide and tetrapeptide segments which inhibited polymerization but not ligand interactions were the amino terminal tripeptide (lacking dolaproine and dolaphenine) and the carboxyl terminal tetrapeptide (lacking dolavaline). We speculate that strong inhibition of other ligand interactions with tubulin requires stable peptide binding to tubulin (i.e. slow dissociation), but that inhibition of polymerization requires only rapid binding to tubulin.

Original languageEnglish (US)
Pages (from-to)1503-1515
Number of pages13
JournalBiochemical Pharmacology
Volume45
Issue number7
DOIs
StatePublished - Apr 6 1993

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dolastatin 10
Tubulin
Cytotoxicity
Isomers
Ligands
Polymerization
Colchicine
Nucleotides
Tubulin Modulators
Vinca Alkaloids
Antimitotic Agents
Leukemia L1210
Peptides
Vinblastine
Mollusca
Valine
Amino Acid Sequence

ASJC Scopus subject areas

  • Pharmacology

Cite this

Differential effects of active isomers, segments, and analogs of dolastatin 10 on ligand interactions with tubulin. Correlation with cytotoxicity. / Bai, Ruoli; Roach, Mary Carmen; Jayaram, Srirangam K.; Barkoczy, Jozef; Pettit, George R.; Luduena, Richard F.; Hamel, Ernest.

In: Biochemical Pharmacology, Vol. 45, No. 7, 06.04.1993, p. 1503-1515.

Research output: Contribution to journalArticle

Bai, Ruoli ; Roach, Mary Carmen ; Jayaram, Srirangam K. ; Barkoczy, Jozef ; Pettit, George R. ; Luduena, Richard F. ; Hamel, Ernest. / Differential effects of active isomers, segments, and analogs of dolastatin 10 on ligand interactions with tubulin. Correlation with cytotoxicity. In: Biochemical Pharmacology. 1993 ; Vol. 45, No. 7. pp. 1503-1515.
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abstract = "Dolastatin 10 is a potent antimitotic peptide isolated from the marine mollusk Dolabella auricularia. Four of its five residues are modified amino acids (in sequence, dolavaline, valine, dolaisoleuine, dolaproine, dolaphenine). Besides inhibiting tubulin polymerization, dolastatin 10 noncompetitively inhibits vinca alkaloid binding to tubulin, inhibits nucleotide exchange and formation of the βs cross-link, and stabilizes the colchicine binding activity of tubulin. To examine the mechanism of action of dolastatin 10 we prepared six chiral isomers, one tri- and one tetrapeptide segment, and one pentapeptide analog of dolastatin 10, all of which differ little from dolastatin 10 as inhibitors of tubulin polymerization. However, only two of the chiral isomers were similar to dolastatin 10 in their cytotoxicity for L1210 murine leukemia cells and in their effects on vinblastine binding, nucleotide exchange, βs cross-link formation, and colchicine binding. These were isomer 2, with reversal of configuration at position C(19a) in the dolaisoleuine moiety, and isomer 19, with reversal of configuration at position C(6) in the dolaphenine moiety. The pentapeptides with reduced cytotoxicity and reduced effects on tubulin interactions with other ligands were all modified in the dolaproine moiety at positions C(9) and/ or C(10). The tripeptide and tetrapeptide segments which inhibited polymerization but not ligand interactions were the amino terminal tripeptide (lacking dolaproine and dolaphenine) and the carboxyl terminal tetrapeptide (lacking dolavaline). We speculate that strong inhibition of other ligand interactions with tubulin requires stable peptide binding to tubulin (i.e. slow dissociation), but that inhibition of polymerization requires only rapid binding to tubulin.",
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