Structure of a RING E3 ligase and ubiquitin-loaded E2 primed for catalysis. / Plechanovová, Anna; Jaffray, Ellis G.; Tatham, Michael H.; Naismith, James H.; Hay, Ronald T.
In: Nature, Vol. 489, No. 7414, 2012, p. 115-120.Research output: Contribution to journal › Article
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TY - JOUR
T1 - Structure of a RING E3 ligase and ubiquitin-loaded E2 primed for catalysis
A1 - Plechanovová,Anna
A1 - Jaffray,Ellis G.
A1 - Tatham,Michael H.
A1 - Naismith,James H.
A1 - Hay,Ronald T.
AU - Plechanovová,Anna
AU - Jaffray,Ellis G.
AU - Tatham,Michael H.
AU - Naismith,James H.
AU - Hay,Ronald T.
PY - 2012
Y1 - 2012
N2 - Ubiquitin modification is mediated by a large family of specificity determining ubiquitin E3 ligases. To facilitate ubiquitin transfer, RING E3 ligases bind both substrate and a ubiquitin E2 conjugating enzyme linked to ubiquitin via a thioester bond, but the mechanism of transfer has remained elusive. Here we report the crystal structure of the dimeric RING domain of rat RNF4 in complex with E2 (UbcH5A) linked by an isopeptide bond to ubiquitin. While the E2 contacts a single protomer of the RING, ubiquitin is folded back onto the E2 by contacts from both RING protomers. The carboxy-terminal tail of ubiquitin is locked into an active site groove on the E2 by an intricate network of interactions, resulting in changes at the E2 active site. This arrangement is primed for catalysis as it can deprotonate the incoming substrate lysine residue and stabilize the consequent tetrahedral transition-state intermediate.
AB - Ubiquitin modification is mediated by a large family of specificity determining ubiquitin E3 ligases. To facilitate ubiquitin transfer, RING E3 ligases bind both substrate and a ubiquitin E2 conjugating enzyme linked to ubiquitin via a thioester bond, but the mechanism of transfer has remained elusive. Here we report the crystal structure of the dimeric RING domain of rat RNF4 in complex with E2 (UbcH5A) linked by an isopeptide bond to ubiquitin. While the E2 contacts a single protomer of the RING, ubiquitin is folded back onto the E2 by contacts from both RING protomers. The carboxy-terminal tail of ubiquitin is locked into an active site groove on the E2 by an intricate network of interactions, resulting in changes at the E2 active site. This arrangement is primed for catalysis as it can deprotonate the incoming substrate lysine residue and stabilize the consequent tetrahedral transition-state intermediate.
U2 - 10.1038/nature11376
DO - 10.1038/nature11376
M1 - Article
JO - Nature
JF - Nature
SN - 0028-0836
IS - 7414
VL - 489
SP - 115
EP - 120
ER -