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Koch, V., Reinders, Y., Ho, H., Sickmann, A., Graf, R., (2006) ' Identification and isolation of Dictyostelium microtubule-associated protein interactors by tandem affinity purification. ' Eur J Cell Biol 85 1079-90

 

Abstract:Tandem affinity purification (TAP) is a method originally established in yeast to isolate highly purified protein complexes in a very gentle and efficient way. In this work, we have modified TAP for Dictyostelium applications and have proved it as a useful method to specifically isolate and identify microtubule-associated protein (MAP) complexes. MAPs are known to interact with other proteins to fulfill their complex functions in balancing the dynamic instability of microtubules as well as anchoring microtubules at the cell cortex, controlling mitosis at the centrosome and guiding transport along them. DdEB1 and the Dictyostelium member of the XMAP215 protein family, DdCP224, are known to be part of complexes at the microtubule tips as well as at the centrosome. Employing TAP and mass spectrometry we were able to prove an interaction between EB1 and the DdCP224. Additionally, among other interactions that remain to be confirmed by other methods, an interaction between DdCP224 and a TACC-family protein could be shown for the first time in Dictyostelium and was confirmed by colocalization and co-immunoprecipitation analyses.
Status:	Published	Type:	Journal article	Source:	PUBMED	PubMed ID:	16782229

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	Genes addressed in this paper
	abpB	acpB	apm1	arcB	calA	cepJ	dymA	eb1
Topics in this paper
Protein-Protein Interactions	X	X	X	X	X	X	X	X
Techniques and Reagents						X		X
Synonym						X		X

	Genes addressed in this paper
	efa1B	efa1G	gpbB	hspB	nog1	pdx2	ranA	sahA
Topics in this paper
Protein-Protein Interactions	X	X	X	X	X	X	X	X
Techniques and Reagents
Synonym

	Genes addressed in this paper
	tacc	talA	tubB	twfA
Topics in this paper
Protein-Protein Interactions	X	X	X	X
Techniques and Reagents
Synonym