TY - JOUR
T1 - Protein complexes characterization in arabidopsis thaliana by tandem affinity purification coupled to mass spectrometry analysis
AU - Bigeard, Jean
AU - Pflieger, Delphine
AU - Colcombet, Jean
AU - Gérard, Loïc
AU - Mireau, Hakim
AU - Hirt, Heribert
N1 - Publisher Copyright:
© Springer Science+Business Media New York 2014.
PY - 2014
Y1 - 2014
N2 - Proteins are major elements participating in all the key functions of the cells. They rarely fulfill their physiological roles in an autonomous way but rather act as part of more complex cellular machines. Indeed they can bind different types of molecules (proteins, nucleic acids, metabolites, etc.), via stable or transient interactions, depending on their nature and functions. The identification of the molecular partners of a given protein is hence essential to better understand its roles, regulation, and mechanisms of action. This chapter describes the use of a tandem affinity purification approach followed by mass spectrometry analysis to try to identify and characterize the proteins involved in protein complexes in Arabidopsis thaliana and decipher some mechanisms of regulation of the modules. Important elements to consider in such an approach are first extensively exposed in the introduction. This technique, in combination with complementary approaches like yeast two-hybrid and bimolecular fluorescence complementation, can be an interesting source of data to identify and characterize in vivo protein complexes.
AB - Proteins are major elements participating in all the key functions of the cells. They rarely fulfill their physiological roles in an autonomous way but rather act as part of more complex cellular machines. Indeed they can bind different types of molecules (proteins, nucleic acids, metabolites, etc.), via stable or transient interactions, depending on their nature and functions. The identification of the molecular partners of a given protein is hence essential to better understand its roles, regulation, and mechanisms of action. This chapter describes the use of a tandem affinity purification approach followed by mass spectrometry analysis to try to identify and characterize the proteins involved in protein complexes in Arabidopsis thaliana and decipher some mechanisms of regulation of the modules. Important elements to consider in such an approach are first extensively exposed in the introduction. This technique, in combination with complementary approaches like yeast two-hybrid and bimolecular fluorescence complementation, can be an interesting source of data to identify and characterize in vivo protein complexes.
KW - Arabidopsis thaliana
KW - Mass spectrometry
KW - Posttranslational modifications
KW - Protein complexes
KW - Tandem affinity purification
UR - http://www.scopus.com/inward/record.url?scp=84922481433&partnerID=8YFLogxK
U2 - 10.1007/978-1-4939-0922-3_18
DO - 10.1007/978-1-4939-0922-3_18
M3 - Article
C2 - 24908132
AN - SCOPUS:84922481433
SN - 1064-3745
VL - 1171
SP - 237
EP - 250
JO - Methods in Molecular Biology
JF - Methods in Molecular Biology
ER -