Site-Specific Identification of SUMO-2 Targets in Cells Reveals an Inverted SUMOylation Motif and a Hydrophobic Cluster SUMOylation Motif

被引:253
作者
Matic, Ivan [1 ]
Schimmel, Joost [2 ]
Hendriks, Ivo A. [2 ]
van Santen, Maria A. [2 ]
van de Rijke, Frans [2 ]
van Dam, Hans [2 ]
Gnad, Florian [1 ]
Mann, Matthias [1 ]
Vertegaal, Alfred C. O. [2 ]
机构
[1] Max Planck Inst Biochem, Dept Prote & Signal Transduct, D-82152 Martinsried, Germany
[2] Leiden Univ, Med Ctr, Dept Mol Cell Biol, NL-2300 RC Leiden, Netherlands
关键词
IN-VIVO IDENTIFICATION; MASS-SPECTROMETRY; QUANTITATIVE PROTEOMICS; PROTEIN SUMOYLATION; PROXIMITY LIGATION; UBIQUITIN; TRANSCRIPTION; CONJUGATION; COMPLEXES; RANGAP1;
D O I
10.1016/j.molcel.2010.07.026
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Reversible protein modification by small ubiquitin-like modifiers (SUMOs) is critical for eukaryotic life. Mass spectrometry-based proteomics has proven effective at identifying hundreds of potential SUMO target proteins. However, direct identification of SUMO acceptor lysines in complex samples by mass spectrometry is still very challenging. We have developed a generic method for the identification of SUMO acceptor lysines in target proteins. We have identified 103 SUMO-2 acceptor lysines in endogenous target proteins. Of these acceptor lysines, 76 are situated in the SUMOylation consensus site [VILMFPC]KxE. Interestingly, eight sites. fit the inverted SUMOylation consensus motif [ED]xK[VILFP]. In addition, we found direct mass spectrometric evidence for crosstalk between SUMOylation and phosphorylation with a preferred spacer between the SUMOylated lysine and the phosphorylated serine of four residues. In 16 proteins we identified a hydrophobic cluster SUMOylation motif (HCSM). SUMO conjugation of RanGAP1 and ZBTB1 via HCSMs is remarkably efficient.
引用
收藏
页码:641 / 652
页数:12
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