Exposure of beta-tubulin regions defined by antibodies on an Arabidopsis thaliana microtubule protofilament model and in the cells.docVIP
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Exposure of beta-tubulin regions defined by antibodies on an Arabidopsis thaliana microtubule protofilament model and in the cells
Blumeetal.BMCPlantBiology2010,10:29
/1471-2229/10/29
RESEARCH ARTICLE
OpenAccess
Exposureofbeta-tubulinregionsdefinedby
antibodiesonanArabidopsisthalianamicrotubule
protofilamentmodelandinthecells
YaroslavBlume1?,AllaYemets1?,YarinaSheremet1,AlexeyNyporko1,VadymSulimenko2,TetyanaSulimenko2,
PavelDráber2*
Abstract
Background:Thefunctionofthecorticalmicrotubules,composedofab-tubulinheterodimers,islinkedtotheir
organizationalstatewhichissubjecttospatialandtemporalmodulationbyenvironmentalcues.Theroleof
tubulinposttranslationalmodificationsintheseprocessesislargelyunknown.Althoughantibodiesagainstsmall
tubulinregionsrepresentusefultoolforstudyingmolecularconfigurationofmicrotubules,dataontheexposureof
tubulinepitopesonplantmicrotubulesarestilllimited.
Results:UsinghomologymodelingwehavegeneratedanArabidopsisthalianamicrotubuleprotofilamentmodel
thatservedforthepredictionofsurfaceexposureoffiveb-tubulinepitopesaswellastyrosineresidues.Peptide
scansnewlydisclosedthepositionofepitopesdetectedbyantibodies18D6(b1-10),TUB2.1(b426-435)andTU-14
(b436-445).Experimentalverificationoftheresultsbyimmunofluorescencemicroscopyrevealedthattheexposure
ofepitopesdependedonthemodeoffixation.Moreover,homologymodelingshowedthatonlytyrosinesinthe
C-terminalregionofb-tubulins(behindb425)wereexposedonthemicrotubuleexternalside.
Immunofluorescencemicroscopyrevealedtyrosinephosphorylationofmicrotubulesinplantcells,implyingthat
b-tubulinscouldbeoneofthetargetsfortyrosinekinases.
Conclusions:Wepredictedsurfaceexposureoffiveb-tubulinepitopes,aswellastyrosineresidues,onthesurface
ofA.thalianamicrotubuleprotofilamentmodel,andvalidatedtheobtainedresultsbyimmunofluorescence
microscopyoncorticalmicrotubulesincells.
Theresultssuggestthatpredictionofepitopeexposureonmicrotubulesbymeansofhomologymodelingcom-
binedwithsite-directedantibodiescancontributetoabetterunderstandingoftheinteractionsofplantmicrotu-
buleswithassociatedproteins.
Background
fashiontoform13protofilamentsthatconstitutecylind-
Microt
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