[25] Microtubules, tubulin, and microtubule-associated proteins of trypanosomes

Robinson, Derrick R.; Beattie, Pauline; Sherwin, Trevor; Gull, Keith

doi:10.1016/0076-6879(91)96027-o

Cited by 84 publications

(76 citation statements)

References 28 publications

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“…4, lane P1). As expected (30), nearly all cellular tubulin remains polymerized and therefore sediments at 1000 ϫ g (Fig. 4, lane P1), demonstrating the resistance of the microtubule-based cytoskeleton to this treatment.…”

Section: Tltf Is a 54-kda Protein That Is Expressed In Both Insectmentioning

confidence: 74%

“…Detergent-extracted cytoskeletons and Ca 2ϩ -resistant flagellar fractions (Fig. 4) were obtained according to the method of Robinson and colleagues (30). All fractions were monitored by phasecontrast microscopy to ensure that lysis and detergent/CaCl 2 extraction was complete.…”

Section: Methodsmentioning

confidence: 99%

“…Whole cells were settled directly onto chilled poly-L-lysine coated coverslips. Cytoskeletons and flagella were prepared in solution (30), prior to being settled onto chilled poly-L-lysine-coated coverslips. All coverslips were washed with PBS to remove unattached material, fixed as indicated in Table I, and either rehydrated (for methanol/acetone fixations) or quenched (for aldehyde fixation) using standard procedures (31).…”

Section: Methodsmentioning

confidence: 99%

“…4, lane S1), indicating that this protein interacts with the membrane in the hypotonic P1 pellet (Fig. 3E), whereas TLTF interacts with the -resistant "flagellar fraction" can then be separated from the solubilized subpellicular cytoskeleton by centrifugation (30).…”

Section: Tltf Is a 54-kda Protein That Is Expressed In Both Insectmentioning

confidence: 99%

“…3, A-C) as has been reported previously (26). Since trypanosome microtubules and associated proteins are highly resistant to physical and chemical disruption (30), they are expected to sediment under these conditions. Hence, the major protein of about 55 kDa seen in the Coomassiestained P1 sample (Fig.…”

Section: Tltf Is a 54-kda Protein That Is Expressed In Both Insectmentioning

confidence: 99%

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T Lymphocyte-triggering Factor of African Trypanosomes Is Associated with the Flagellar Fraction of the Cytoskeleton and Represents a New Family of Proteins That Are Present in Several Divergent Eukaryotes

Hill¹,

Hutchings²,

Grandgenett³

et al. 2000

Journal of Biological Chemistry

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Section: Tltf Is a 54-kda Protein That Is Expressed In Both Insectmentioning

confidence: 74%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Tltf Is a 54-kda Protein That Is Expressed In Both Insectmentioning

confidence: 99%

Section: Tltf Is a 54-kda Protein That Is Expressed In Both Insectmentioning

confidence: 99%

See 3 more Smart Citations

T Lymphocyte-triggering Factor of African Trypanosomes Is Associated with the Flagellar Fraction of the Cytoskeleton and Represents a New Family of Proteins That Are Present in Several Divergent Eukaryotes

Hill¹,

Hutchings²,

Grandgenett³

et al. 2000

Journal of Biological Chemistry

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Selective and reversible effects of vinca alkaloids onTrypanosoma cruzi epimastigote forms: Blockage of cytokinesis without inhibition of the organelle duplication

Grellier

Sinou

Loubresse

et al. 1999

Cell Motil. Cytoskeleton

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In vivo and in vitro phosphorylation and subcellular localization of trypanosomatid cytoskeletal giant proteins

Baqui

Milder

Mortara

et al. 2000

Cell Motil. Cytoskeleton

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Polyclonal antibodies to Lt 1200 and to Ps 3500 specifically recognize similar polypeptides of the same genera of parasite. In addition to reacting with giant polypeptides of the Leptomonas species, anti-Ls 2500 also cross reacts with Ps 3500, and with a 500-kDa polypeptide of Leishmania. Confocal immunofluorescence and immunogold electron microscopy showed major differences in topological distribution of these three proteins, though they partially share a common localization at the anterior end of the cell body skeleton. Furthermore, Ps 3500, Ls 2500, and Lt 1200 are in vivo phosphorylated at serine and threonine residues, whereas, in vitro phosphorylation of cytoskeletal fractions reveal that only Ps 3500 and Ls 2500 are phosphorylated. Heat treatment (100°C) of high salt cytoskeletal extracts demonstrates that Ps 3500 and Ls 2500 remain stable in solution, whereas Lt 1200 is denatured. Kinase assays with immunocomplexes of heat-treated giant proteins show that only Ps 3500 and Ls 2500 are phosphorylated. These results demonstrate the existence of a novel class of megadalton phosphoproteins in promastigote forms of trypanosomatids that appear to be genera specific with distinct cytoskeletal functions. In addition, there is also evidence that Ps 3500 and Ls 2500, in contrast to Lt 1200, seem to be autophosphorylating serine and threonine protein kinases, suggesting that they might play regulatory roles in the cytoskeletal organization.

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[25] Microtubules, tubulin, and microtubule-associated proteins of trypanosomes

Cited by 84 publications

References 28 publications

T Lymphocyte-triggering Factor of African Trypanosomes Is Associated with the Flagellar Fraction of the Cytoskeleton and Represents a New Family of Proteins That Are Present in Several Divergent Eukaryotes

T Lymphocyte-triggering Factor of African Trypanosomes Is Associated with the Flagellar Fraction of the Cytoskeleton and Represents a New Family of Proteins That Are Present in Several Divergent Eukaryotes

Selective and reversible effects of vinca alkaloids onTrypanosoma cruzi epimastigote forms: Blockage of cytokinesis without inhibition of the organelle duplication

In vivo and in vitro phosphorylation and subcellular localization of trypanosomatid cytoskeletal giant proteins

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