UCH-L5 in complex with ubiquitin-propargyl bound to the RPN13 DEUBAD domain

Mechanism of Uch-L5 Activation and Inhibition by Deubad Domains in Rpn13 and Ino80G.

ACTIVE SITE CYS88 COVALENTLY LINKED TO UBIQUITIN-PROPARGYL
UBIQUITIN CARBOXYL-TERMINAL HYDROLASE ISOZYME L5

GLY76 OF UBIQUITIN IS REPLACED BY GLY-AYE THAT IS COVALENTLY LINKED TO THE ACTIVE SITE CYS88 OF UCH-L5

UCH-L5, UBIQUITIN C-TERMINAL HYDROLASE UCH37, UBIQUITIN THIOESTERASE L5

RPN13,110 KDA CELL MEMBRANE GLYCOPROTEIN, GP110, ADHESION-REGULATING MOLECULE 1, ARM-1, PROTEASOME REGULATORY PARTICLE NON-ATPASE 13, HRPN13, RPN13 HOMOLOG

GPGMTGNAGEWCLMESDPGVFTELIKGFGCRGAQVEEIWSLEPENFEKLKPVHGLIFLFKWQPGEEPAGSVVQDSRLDTI
FFAKQVINNACATQAIVSVLLNCTHQDVHLGETLSEFKEFSQSFDAAMKGLALSNSDVIRQVHNSFARQQMFEFDTKTSA
KEEDAFHFVSYVPVNGRLYELDGLREGPIDLGACNQDDWISAVRPVIEKRIQKYSEGEIRFNLMAIVSDRKMIYEQKIAE
LQRQLAEEPMDTDQGNSMLSAIQSEVAKNQMLIEEEVQKLKRYKIENIRRKHNYLPFIMELLKTLAEHQQLIPLVEKAKE
KQNAKKAQETK

GPGSDLQSILATMNVPAGPAGGQQVDLASVLTPEIMAPILANADVQERLLPYLPSGESLPQTADEIQNTLTSPQFQQALG
MFSAALASGQLGPLMCQFGLPAEAVEAANKGDVEAFAKAMQNNAKPE

MOLECULAR REPLACEMENT WAS PERFORMED WITH THE PARTIALLY BUILD UCH-L5-RPN13 STRUCTURE, SEE RELATED PDB

100 MM BIS-TRIS-PROPANE PH 5.8, 300 MM NABR, 21% PEG3350. 4 DEGREES CELSIUS

UCH-L5 IN COMPLEX WITH THE RPN13 DEUBAD DOMAIN

GLY76 IN NATIVE UBIQUITIN IS REPLACED BY AYE

HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS.

UBIQUITIN CARBOXYL-TERMINAL HYDROLASE ISOZYME L5 (E.C.3.4.19.12), POLYUBIQUITIN-B, PROTEASOMAL UBIQUITIN RECEPTOR ADRM1

HYDROLASE, DEUBIQUITINATING ENZYME, DUB, UCH37, UCHL5, PROTEASOME, ADRM1, RPN13, DEUBAD, UCH, UBIQUITIN-PROPARGYL