Literature summary for 6.2.1.45 extracted from

Oweis, W.; Padala, P.; Hassouna, F.; Cohen-Kfir, E.; Gibbs, D.R.; Todd, E.A.; Berndsen, C.E.; Wiener, R.
Trans-binding mechanism of ubiquitin-like protein activation revealed by a UBA5-UFM1 complex (2016), Cell Rep., 16, 3113-3120 .

Search for more literature for 6.2.1.45

Cloned(Commentary)

Cloned (Comment)	Organism
recombinant expression of N-terminally His6-tagged enzyme UBA5 in Escherichia coli	Homo sapiens

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified UBA5-UFM1 complex, containing both the adenylation domain and the UIS of UBA5, X-ray diffraction structure determination and analysis at 1.85-2.10 A	Homo sapiens

Protein Variants

Protein Variants	Comment	Organism
D290K/C250A	in this heterodimer, the UBA5 subunit that can form the thioester bond with UFM1 is missing the UFC1 binding site. In the UBA5 (D290K)-UBA5 (K271D/C250A DELTADUIS) heterodimer, binding to the UIS and charging can only take place on the same monomer, thereby supporting a cis-binding mechanism	Homo sapiens

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	required	Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
ATP + ubiquitin + [E1 ubiquitin-activating enzyme]-L-cysteine	Homo sapiens	-	AMP + diphosphate + S-ubiquitinyl-[E1 ubiquitin-activating enzyme]-L-cysteine	-	?

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	Q9GZZ9	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant N-terminally His6-tagged enzyme UBA5 from Escherichia coli by nickel affinity chromatography	Homo sapiens

Reaction

Reaction	Comment	Organism	Reaction ID
ATP + ubiquitin + [E1 ubiquitin-activating enzyme]-L-cysteine = AMP + diphosphate + S-ubiquitinyl-[E1 ubiquitin-activating enzyme]-L-cysteine	modeling of a cis-binding mechanism of UFM1 to UBA5, trans-binding mechanism of UFM1 transfer to the E2, UFC1, and UFM1-UBA5 adenylation domain interactions, overview. Although the active site Cys in the UBA5 apo structure is located at the N terminus of helix H, this Cys is no longer part of a helical structure in the complex with UFM1. Conformational changes in UBA5 are required to bring the active site Cys into the vicinity of UFM1-AMP mixed anhydride bond, leading to formation of the thioester bond	Homo sapiens	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
ATP + ubiquitin + [E1 ubiquitin-activating enzyme]-L-cysteine	-	Homo sapiens	AMP + diphosphate + S-ubiquitinyl-[E1 ubiquitin-activating enzyme]-L-cysteine	-	?
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	?	-	?

Subunits

Subunits	Comment	Organism
homodimer	dimerization of UBA5 is required for UFM1 activation. The active site Cys of UBA5 (Cys 250) is located within the adenylation domain	Homo sapiens

Synonyms

Synonyms	Comment	Organism
UBA5	-	Homo sapiens

Cofactor

Cofactor	Comment	Organism	Structure
ATP	-	Homo sapiens

General Information

General Information	Comment	Organism
additional information	UBA5 is the smallest and structurally simplest E1. The active site Cys of UBA5 (Cys 250) is located within the adenylation domain, but this domain is not sufficient for the formation of a thioester bond between the UFM1 C terminus and the UBA5 catalytic Cys. Modeling of a cis-binding mechanism of UFM1 to UBA5. Trans-binding mechanism of UFM1 transfer to the E2, UFC1. Homodimerization of UBA5 is essential for activating UFM1	Homo sapiens

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Release 2023.1 (January 2023)