6.2.1.45 ATP + Oregon Green-labeled ubiquitin + [ubiquitin-activating protein E1]-L-cysteine enzyme efficiently accepts ubiquitin substrate fluorescently labeled by Oregon Green Homo sapiens AMP + diphosphate + [ubiquitin-activating protein E1]-S-(Oregon Green-labeled ubiquitinyl)-L-cysteine - ? 428389 6.2.1.45 ATP + SUMO2 + [ubiquitin-activating protein UBA5]-L-cysteine SUMO2, small ubiquitin-like modifier2, an ubiquitin-like protein Homo sapiens AMP + diphosphate + [ubiquitin-activating protein UBA5]-S-SUMO2-L-cysteine enzyme greatly activates SUMO2 in the nucleus or transfers activated SUMO2 to the nucleus after it conjugated SUMO2 in the cytoplasm ? 428399 6.2.1.45 ATP + ubiquitin + SUMO2 UBE1DC1 greatly activates SUMO2 in the nucleus or transfers activated-SUMO2 to nucleus after conjugation of SUMO2 in the cytoplasm Homo sapiens ? - ? 400478 6.2.1.45 ATP + ubiquitin + ubiquitin carrier protein E2 - Homo sapiens AMP + diphosphate + ubiquitin-(ubiquitin carrier protein E2) - ? 502 6.2.1.45 ATP + ubiquitin + ubiquitin-fold modifier 1 - Homo sapiens ? - ? 400498 6.2.1.45 ATP + ubiquitin + Ufm1 - Homo sapiens ? - ? 400499 6.2.1.45 ATP + ubiquitin + [6His-ubiquitin-activating enzyme E1]W-8His-Strep-HA Strep, i.e.WSHPQFEK, HA, i.e. YPYDVPDYAS, under non-reducing conditions, the intermediate complex of the thioester formation is not observed without ATP Homo sapiens AMP + diphosphate + [6His-ubiquitin-activating enzyme E1]W-8His-Strep-HA-ubiquitinyl-L-cysteine - ? 425986 6.2.1.45 ATP + ubiquitin + [E1 ubiquitin-activating enzyme]-L-cysteine - Drosophila melanogaster AMP + diphosphate + S-ubiquitinyl-[E1 ubiquitin-activating enzyme]-L-cysteine - ? 432709 6.2.1.45 ATP + ubiquitin + [E1 ubiquitin-activating enzyme]-L-cysteine - Mus musculus AMP + diphosphate + S-ubiquitinyl-[E1 ubiquitin-activating enzyme]-L-cysteine - ? 432709 6.2.1.45 ATP + ubiquitin + [E1 ubiquitin-activating enzyme]-L-cysteine - Homo sapiens AMP + diphosphate + S-ubiquitinyl-[E1 ubiquitin-activating enzyme]-L-cysteine - ? 432709 6.2.1.45 ATP + ubiquitin + [E1 ubiquitin-activating enzyme]-L-cysteine - Schizosaccharomyces pombe AMP + diphosphate + S-ubiquitinyl-[E1 ubiquitin-activating enzyme]-L-cysteine - ? 432709 6.2.1.45 ATP + ubiquitin + [E1 ubiquitin-activating enzyme]-L-cysteine - Citrus reticulata AMP + diphosphate + S-ubiquitinyl-[E1 ubiquitin-activating enzyme]-L-cysteine - ? 432709 6.2.1.45 ATP + ubiquitin + [E1 ubiquitin-activating enzyme]-L-cysteine - Arabidopsis thaliana AMP + diphosphate + S-ubiquitinyl-[E1 ubiquitin-activating enzyme]-L-cysteine - ? 432709 6.2.1.45 ATP + ubiquitin + [E1 ubiquitin-activating enzyme]-L-cysteine - Giardia intestinalis AMP + diphosphate + S-ubiquitinyl-[E1 ubiquitin-activating enzyme]-L-cysteine - ? 432709 6.2.1.45 ATP + ubiquitin + [E1 ubiquitin-activating enzyme]-L-cysteine - Ovis aries AMP + diphosphate + S-ubiquitinyl-[E1 ubiquitin-activating enzyme]-L-cysteine - ? 432709 6.2.1.45 ATP + ubiquitin + [E1 ubiquitin-activating enzyme]-L-cysteine - Saccharomyces cerevisiae AMP + diphosphate + S-ubiquitinyl-[E1 ubiquitin-activating enzyme]-L-cysteine - ? 432709 6.2.1.45 ATP + ubiquitin + [E1 ubiquitin-activating enzyme]-L-cysteine E1-activating enzyme activates ubiquitin via an adenylated intermediate and catalyzes its transfer to an E2 enzyme Homo sapiens AMP + diphosphate + S-ubiquitinyl-[E1 ubiquitin-activating enzyme]-L-cysteine - ? 432709 6.2.1.45 ATP + ubiquitin + [E1 ubiquitin-activating enzyme]-L-cysteine E1 activates ubiquitin or an ubiquitin-like protein and transfers it to the E2-conjugating enzyme Homo sapiens AMP + diphosphate + S-ubiquitinyl-[E1 ubiquitin-activating enzyme]-L-cysteine - ? 432709 6.2.1.45 ATP + ubiquitin + [E1 ubiquitin-activating enzyme]-L-cysteine - Saccharomyces cerevisiae ATCC 204508 AMP + diphosphate + S-ubiquitinyl-[E1 ubiquitin-activating enzyme]-L-cysteine - ? 432709 6.2.1.45 ATP + ubiquitin + [E1 ubiquitin-activating enzyme]-L-cysteine - Giardia intestinalis ATCC 50803 AMP + diphosphate + S-ubiquitinyl-[E1 ubiquitin-activating enzyme]-L-cysteine - ? 432709 6.2.1.45 ATP + ubiquitin + [ubiquitin-activating enzyme Uba5]-L-cysteine the catalytic cysteine residue of isoform Uba5 is part of the adenylation domain in a alpha-helical motif Homo sapiens AMP + diphosphate + [ubiquitin-activating enzyme Uba5]-S-ubiquitinyl-L-cysteine - ? 428402 6.2.1.45 ATP + ubiquitin + [ubiquitin-activating protein E1]-L-cysteine - Mus musculus AMP + diphosphate + [ubiquitin-activating protein E1]-S-ubiquitinyl-L-cysteine - ? 425987 6.2.1.45 ATP + ubiquitin + [ubiquitin-activating protein E1]-L-cysteine - Homo sapiens AMP + diphosphate + [ubiquitin-activating protein E1]-S-ubiquitinyl-L-cysteine - ? 425987 6.2.1.45 ATP + ubiquitin + [ubiquitin-activating protein E1]-L-cysteine - Bos taurus AMP + diphosphate + [ubiquitin-activating protein E1]-S-ubiquitinyl-L-cysteine - ? 425987 6.2.1.45 ATP + ubiquitin + [ubiquitin-activating protein E1]-L-cysteine a carboxylgroup is first activated as an adenylate followed by its direct transfer to an autonomous molecular moiety in a single enzymatic step Escherichia coli AMP + diphosphate + [ubiquitin-activating protein E1]-S-ubiquitinyl-L-cysteine - ? 425987 6.2.1.45 ATP + ubiquitin + [ubiquitin-activating protein Uba1a]-L-cysteine - Homo sapiens AMP + diphosphate + [ubiquitin-activating protein Uba1a]-S-ubiquitinyl-L-cysteine - ? 428404 6.2.1.45 ATP + ubiquitin + [ubiquitin-activating protein UBA1]-L-cysteine enzyme forms higher molecular mass intermediates with ubiquitin Arabidopsis thaliana AMP + diphosphate + [ubiquitin-activating protein UBA1]-S-ubiquitinyl-L-cysteine the enzyme-ubiquitin intermediates dissociate in presence of 2-mercaptoethanol, indicating thiolester linkage ? 428405 6.2.1.45 ATP + ubiquitin + [ubiquitin-activating protein UBA2]-L-cysteine enzyme forms higher molecular mass intermediates with ubiquitin Arabidopsis thaliana AMP + diphosphate + [ubiquitin-activating protein UBA2]-S-ubiquitinyl-L-cysteine the enzyme-ubiquitin intermediates dissociate in presence of 2-mercaptoethanol, indicating thiolester linkage ? 428406 6.2.1.45 ATP + ubiquitin + [ubiquitin-activating protein Uba6]-L-cysteine isoform Uba6 forms a covalent link with ubiquitin in vitro and in vivo, which is sensitive to reducing conditions. Recombinant E1 enzyme Uba6 can activate ubiquitin and transfer it onto the ubiquitin-conjugating enzyme UbcH5B. Ubiquitin activated by Uba6 can be used for ubiquitylation of p53 and supports the autoubiquitylation of the E3 ubiquitin ligases HectH9 and E6-AP Homo sapiens AMP + diphosphate + [ubiquitin-activating protein Uba6]-S-ubiquitinyl-L-cysteine - ? 428408 6.2.1.45 ATP + ubiquitin + [ubiquitin-activating protein UBE1]-L-cysteine - Homo sapiens AMP + diphosphate + [ubiquitin-activating protein UBE1]-S-ubiquitinyl-L-cysteine - ? 428409 6.2.1.45 ATP + ubiquitin fold modifier1 + [ubiquitin-activating enzyme Uba5]-L-cysteine the catalytic cysteine residue of isoform Uba5 is part of the adenylation domain in a alpha-helical motif Homo sapiens AMP + diphosphate + [ubiquitin-activating enzyme Uba5]-S-(ubiquitin fold modifier1)-L-cysteine - ? 428410 6.2.1.45 ATP + ubiquitin mutant G76A + [ubiquitin-activating protein E1]-L-cysteine - Bos taurus AMP + diphosphate + [ubiquitin-activating protein E1]-S-(ubiquitin mutant G76A)yl-L-cysteine mutant ubiquitin G76A, bearing a Gly to Ala substitution at the COOH terminus is a substrate for El enzyme. Ubiquitin G76A supports PPI-ATP exchange with 500fold decrease in kcat/Km compared to wild-type ubiquitin, does not produce detectable AMP-Ub with native El, produces stoichiometric AMP-Ub with thiol-blocked El, gives a stoichiometric burst of ATP consumption with either native or thiol-blocked El, support El-ubiquitin thiol ester formation with native El, and supports several downstream reactions of the proteolytic pathway with a 20% decrease to the rate of wild type ubiquitin ? 428411 6.2.1.45 ATP + Ufm1 + [ubiquitin-activating protein UBA5]-L-cysteine Ufm1, ubiquitin-fold modifier 1, an ubiquitin-like protein Homo sapiens AMP + diphosphate + [ubiquitin-activating protein UBA5]-S-Ufm1-L-cysteine - ? 428417 6.2.1.45 additional information impaired nucleotide excision repair upon macrophage differentiation is corrected by E1 ubiquitin-activating enzyme Homo sapiens ? - ? 89 6.2.1.45 additional information UBE1L2 transfers activated ubiquitin onto UbcH5b and supports E3-mediated polyubiquitylation Homo sapiens ? - ? 89 6.2.1.45 additional information a lysine 48-linked polyubiquitin chain, assembled upon an internal lysine residue of a substrate protein, becomes the principle signal for recognition and target degradation by the 26S proteasome. E1 is not only essential for the initial ATP-dependent activation of ubiquitin in the ubiquitin degradtion pathway, but also capable of the catalytic extension of the polyubiquitin chain on a mono-ubiquitinated substrate Escherichia coli ? - ? 89 6.2.1.45 additional information E1 consumes ATP and converts ubiquitin to a transfer-competent, enzyme-bound thioester. The reaction begins with ubiquitin-adenylate formation and the release of diphosohate. The active site cysteine of the E1 then displaces the AMP leading to a ubiquitin-E1 thioester complex Mus musculus ? - ? 89 6.2.1.45 additional information E1 activity is assesssed by the capacity of the enzyme to form a thiol ester conjugate with ubiquitin in an ATP-dependent process and to transfer this activated ubiquitin molecule to an conjugating enzyme Mus musculus ? - ? 89 6.2.1.45 additional information the thioester formation assay is performed using recombinant proteins expressed in Escherichia coli. The activation of ubiquitin by purified UBE1 is identified in vitro by SDS-PAGE Homo sapiens ? - ? 89 6.2.1.45 additional information kinetics for Uba1a-catalyzed transthiolation of Ubc2b are used as a reporter assay for determining the Km and kcat values for the three cosubstrates of the ubiquitin-activating enzyme. The E2 transthiolation assays are more sensitive to the potential presence of trace catalytically active fragments than the single turnover end point assays used for quantitating ternary complex stoichiometry Homo sapiens ? - ? 89 6.2.1.45 additional information purified isoform UBE1 can activate and conjugate ubiquitin to ubiquitin-conjugating enzyme E2s. Transfer is restricted to distinct E2 isoforms UB2R2, UBE2W and UBE2NL Homo sapiens ? - ? 89 6.2.1.45 additional information residue Cys194 lies within a region of identity to active-site Cys88 of the ubiquitin carrier protein E2, suggesting a potential role for this region in enzymatic function. Residue Cys454 lies within a region of identity to the thiol ester consensus sequence of several proteins involved in thioester formation Homo sapiens ? - ? 89 6.2.1.45 additional information chimeric mutant Aos1-Uba2 SUMO-E1 enzyme shows SUMO-E1 activity. The E1 enzyme catalyzes the formation of a thioester-linked complex between SUMO and the E2 enzyme. This process is initiated by activation of the carboxyl terminus of SUMO by adenylation, followed by a thioesterification reaction in which SUMO is conjugated to a cysteine residue at the active site of Uba2 in the E1 enzyme. SUMO is then transferred to the active site cysteine of the E2 enzyme, Ubc9, via a trans-thioesterification reaction. A SUMO-charged E2 enzyme and substrate are finally bound with or without the assistance of a distinct class of SUMO E3-ligases, resulting in the activated SUMO bound to the substrate through an isopeptide linkage Mus musculus ? - ? 89 6.2.1.45 additional information E1 ubiquitin-activating enzyme UBA6 is the only E1 enzyme that can activate both ubiquitin and ubiquitin-like protein HLA-F adjacent transcript 10 (FAT10). FAT10 consists of two ubiquitin-like domains with 29% and 36% identity to ubiquitin, respectively, that are separated by a short linker region Homo sapiens ? - ? 89 6.2.1.45 additional information orthogonal ubiquitin transfer (OUT) technology to profile their ubiquitination targets in mammalian cells of isozymes Uba1 and Uba6 Homo sapiens ? - ? 89 6.2.1.45 additional information CDC42 is a substrate of UBA6-initiated ubiquitination Homo sapiens ? - ? 89 6.2.1.45 additional information the non-canonical E1, UBA5, binds to the ubiquitin-like protein UFM1 using a trans-binding mechanism in which UFM1 interacts with distinct sites in both subunits of the UBA5 dimer. Mechanism of UFM1 activation by UBA5 and trans-binding mechanism of UFM1 transfer to the E2, UFC1. UFM1 contains a C-terminal Val-Gly dipeptide instead of the canonical Gly-Gly dipeptide present in ubiquitin and other ubiquitin-like proteins Homo sapiens ? - ? 89