TY - JOUR
T1 - A proteomic study of SUMO-2 target proteins
AU - Vertegaal, Alfred C.O.
AU - Ogg, Stephen C.
AU - Jaffray, Ellis
AU - Rodriguez, Manuel S.
AU - Hay, Ronald T.
AU - Andersen, Jens S.
AU - Mann, Matthias
AU - Lamond, Angus I.
PY - 2004/8/6
Y1 - 2004/8/6
N2 - The SUMO family in vertebrates includes at least three distinct proteins (SUMO-1, -2, and -3) that are added as post-translational modifications to target proteins. A considerable number of SUMO-1 target proteins have been identified, but little is known about SUMO-2. A stable HeLa cell line expressing His6-tagged SUMO-2 was established and used to label and purify novel endogenous SUMO-2 target proteins. Tagged forms of SUMO-2 were functional and localized predominantly in the nucleus. His6-tagged SUMO-2 conjugates were affinity-purified from nuclear fractions and identified by mass spectrometry. Eight novel potential SUMO-2 target proteins were identified by at least two peptides. Three of these proteins, SART1, heterogeneous nuclear ribonucleoprotein (RNP) M, and the U5 small nuclear RNP 200-kDa helicase, play a role in RNA metabolism. SART1 and heterogeneous nuclear RNP M were both shown to be genuine SUMO targets, confirming the validity of the approach.
AB - The SUMO family in vertebrates includes at least three distinct proteins (SUMO-1, -2, and -3) that are added as post-translational modifications to target proteins. A considerable number of SUMO-1 target proteins have been identified, but little is known about SUMO-2. A stable HeLa cell line expressing His6-tagged SUMO-2 was established and used to label and purify novel endogenous SUMO-2 target proteins. Tagged forms of SUMO-2 were functional and localized predominantly in the nucleus. His6-tagged SUMO-2 conjugates were affinity-purified from nuclear fractions and identified by mass spectrometry. Eight novel potential SUMO-2 target proteins were identified by at least two peptides. Three of these proteins, SART1, heterogeneous nuclear ribonucleoprotein (RNP) M, and the U5 small nuclear RNP 200-kDa helicase, play a role in RNA metabolism. SART1 and heterogeneous nuclear RNP M were both shown to be genuine SUMO targets, confirming the validity of the approach.
UR - http://www.scopus.com/inward/record.url?scp=4043051557&partnerID=8YFLogxK
U2 - 10.1074/jbc.M404201200
DO - 10.1074/jbc.M404201200
M3 - Article
C2 - 15175327
AN - SCOPUS:4043051557
SN - 0021-9258
VL - 279
SP - 33791
EP - 33798
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 32
ER -