Isolation and mass spectrometry of transcription factor complexes
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Isolation and mass spectrometry of transcription factor complexes. / Sebastiaan Winkler, G.; Lacomis, Lynne; Philip, John; Erdjument-Bromage, Hediye; Svejstrup, Jesper Q.; Tempst, Paul.
I: Methods, Bind 26, Nr. 3, 2002, s. 260-269.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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TY - JOUR
T1 - Isolation and mass spectrometry of transcription factor complexes
AU - Sebastiaan Winkler, G.
AU - Lacomis, Lynne
AU - Philip, John
AU - Erdjument-Bromage, Hediye
AU - Svejstrup, Jesper Q.
AU - Tempst, Paul
N1 - Funding Information: Work in our laboratories was supported by grants from the Imperial Cancer Research Fund and the Human Frontier Science Program Project RG0193/97 (to J.Q.S.) and by NCI Core Grant P30 CA08748 (to P.T.). G.S.W. was supported by an EMBO Long Term Fellowship.
PY - 2002
Y1 - 2002
N2 - Protocols are described that enable the isolation of novel proteins associated with a known protein and the subsequent identification of these proteins by mass spectrometry. We review the basics of nanosample handling and of two complementary approaches to mass analysis, and provide protocols for the entire process. The protein isolation procedure is rapid and based on two high-affinity chromatography steps. The method does not require previous knowledge of complex composition or activity and permits subsequent biochemical characterization of the isolated factor. As an example, we provide the procedures used to isolate and analyze yeast Elongator, a histone acetyltransferase complex important for transcript elongation, which led to the identification of three novel subunits.
AB - Protocols are described that enable the isolation of novel proteins associated with a known protein and the subsequent identification of these proteins by mass spectrometry. We review the basics of nanosample handling and of two complementary approaches to mass analysis, and provide protocols for the entire process. The protein isolation procedure is rapid and based on two high-affinity chromatography steps. The method does not require previous knowledge of complex composition or activity and permits subsequent biochemical characterization of the isolated factor. As an example, we provide the procedures used to isolate and analyze yeast Elongator, a histone acetyltransferase complex important for transcript elongation, which led to the identification of three novel subunits.
UR - http://www.scopus.com/inward/record.url?scp=0036490552&partnerID=8YFLogxK
U2 - 10.1016/S1046-2023(02)00030-0
DO - 10.1016/S1046-2023(02)00030-0
M3 - Journal article
C2 - 12054882
AN - SCOPUS:0036490552
VL - 26
SP - 260
EP - 269
JO - Methods
JF - Methods
SN - 1046-2023
IS - 3
ER -
ID: 331042636