Literature summary for 2.4.3.4 extracted from

Ortiz-Soto, M.E.; Reising, S.; Schlosser, A.; Seibel, J.
Structural and functional role of disulphide bonds and substrate binding residues of the human beta-galactoside alpha-2,3-sialyltransferase 1 (hST3Gal1) (2019), Sci. Rep., 9, 17993 .

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli Origami2 (DE3) cells	Homo sapiens

Protein Variants

Protein Variants	Comment	Organism
C59S/C64S	the mutant shows reduced activity compared to the wild type enzyme	Homo sapiens
H299A	inactive	Homo sapiens
H316A	inactive	Homo sapiens
N147S	the mutant shows strongly reduced activity compared to the wild type enzyme	Homo sapiens
N170A	the mutant shows strongly reduced activity compared to the wild type enzyme	Homo sapiens
S148A	the mutant shows strongly reduced activity compared to the wild type enzyme	Homo sapiens
V315A	the mutant shows strongly reduced activity compared to the wild type enzyme	Homo sapiens
V315Y	the mutant shows strongly reduced activity compared to the wild type enzyme	Homo sapiens
Y191A	the mutant shows strongly reduced activity compared to the wild type enzyme	Homo sapiens
Y230A	the mutant shows strongly reduced activity compared to the wild type enzyme	Homo sapiens
Y230F	the mutant shows strongly reduced activity compared to the wild type enzyme	Homo sapiens
Y300A	inactive	Homo sapiens

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.02	-	benzyl 2-acetamido-2-deoxy-3-O-(beta-D-galactopyranosyl)-alpha-D-galactopyranoside	mutant enzyme C59S/C64S, at pH 7.4 and 37°C	Homo sapiens
0.026	-	benzyl 2-acetamido-2-deoxy-3-O-(beta-D-galactopyranosyl)-alpha-D-galactopyranoside	wild type enzyme, at pH 7.4 and 37°C	Homo sapiens
0.096	-	benzyl 2-acetamido-2-deoxy-3-O-(beta-D-galactopyranosyl)-alpha-D-galactopyranoside	mutant enzyme N170A, at pH 7.4 and 37°C	Homo sapiens
0.105	-	benzyl 2-acetamido-2-deoxy-3-O-(beta-D-galactopyranosyl)-alpha-D-galactopyranoside	mutant enzyme S148A, at pH 7.4 and 37°C	Homo sapiens
0.106	-	CMP-N-acetyl-beta-neuraminate	wild type enzyme, at pH 7.4 and 37°C	Homo sapiens
0.121	-	CMP-N-acetyl-beta-neuraminate	mutant enzyme C59S/C64S, at pH 7.4 and 37°C	Homo sapiens
0.176	-	CMP-N-acetyl-beta-neuraminate	mutant enzyme Y230F, at pH 7.4 and 37°C	Homo sapiens
0.18	-	CMP-N-acetyl-beta-neuraminate	mutant enzyme V315A, at pH 7.4 and 37°C	Homo sapiens
0.184	-	CMP-N-acetyl-beta-neuraminate	mutant enzyme N170A, at pH 7.4 and 37°C	Homo sapiens
0.205	-	CMP-N-acetyl-beta-neuraminate	mutant enzyme Y230A, at pH 7.4 and 37°C	Homo sapiens
0.211	-	benzyl 2-acetamido-2-deoxy-3-O-(beta-D-galactopyranosyl)-alpha-D-galactopyranoside	mutant enzyme N147S, at pH 7.4 and 37°C	Homo sapiens
0.271	-	CMP-N-acetyl-beta-neuraminate	mutant enzyme V315Y, at pH 7.4 and 37°C	Homo sapiens
0.352	-	benzyl 2-acetamido-2-deoxy-3-O-(beta-D-galactopyranosyl)-alpha-D-galactopyranoside	mutant enzyme Y230F, at pH 7.4 and 37°C	Homo sapiens
0.444	-	benzyl 2-acetamido-2-deoxy-3-O-(beta-D-galactopyranosyl)-alpha-D-galactopyranoside	mutant enzyme Y191A, at pH 7.4 and 37°C	Homo sapiens
0.481	-	CMP-N-acetyl-beta-neuraminate	mutant enzyme S148A, at pH 7.4 and 37°C	Homo sapiens
0.549	-	benzyl 2-acetamido-2-deoxy-3-O-(beta-D-galactopyranosyl)-alpha-D-galactopyranoside	mutant enzyme V315Y, at pH 7.4 and 37°C	Homo sapiens
0.668	-	CMP-N-acetyl-beta-neuraminate	mutant enzyme N147S, at pH 7.4 and 37°C	Homo sapiens
0.854	-	benzyl 2-acetamido-2-deoxy-3-O-(beta-D-galactopyranosyl)-alpha-D-galactopyranoside	mutant enzyme Y230A, at pH 7.4 and 37°C	Homo sapiens
0.876	-	CMP-N-acetyl-beta-neuraminate	mutant enzyme Y191A, at pH 7.4 and 37°C	Homo sapiens
1.388	-	benzyl 2-acetamido-2-deoxy-3-O-(beta-D-galactopyranosyl)-alpha-D-galactopyranoside	mutant enzyme V315A, at pH 7.4 and 37°C	Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
CMP-N-acetyl-beta-neuraminate + a beta-D-galactosyl-(1->3)-N-acetyl-alpha-D-galactosaminyl-R	Homo sapiens	-	CMP + an N-acetyl-alpha-neuraminyl-(2->3)-beta-D-galactosyl-(1->3)-N-acetyl-alpha-D-galactosaminyl-R	-	?

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	Q11201	-	-

Purification (Commentary)

Purification (Comment)	Organism
His-Select nickel affinity column chromatography and Superdex S200 gel filtration	Homo sapiens

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
CMP-N-acetyl-beta-neuraminate + a beta-D-galactosyl-(1->3)-N-acetyl-alpha-D-galactosaminyl-R	-	Homo sapiens	CMP + an N-acetyl-alpha-neuraminyl-(2->3)-beta-D-galactosyl-(1->3)-N-acetyl-alpha-D-galactosaminyl-R	-	?
CMP-N-acetyl-beta-neuraminate + asialofetuin	-	Homo sapiens	CMP + ?	-	?
CMP-N-acetyl-beta-neuraminate + benzyl 2-acetamido-2-deoxy-3-O-(beta-D-galactopyranosyl)-alpha-D-galactopyranoside	-	Homo sapiens	CMP + ?	-	?

Subunits

Subunits	Comment	Organism
?	x * 66200, maltose-binding-protein-fused recombinant enzyme, SDS-PAGE	Homo sapiens

Synonyms

Synonyms	Comment	Organism
beta-galactoside alpha-2,3-sialyltransferase 1	-	Homo sapiens
ST3GAL1	-	Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
0.13	-	CMP-N-acetyl-beta-neuraminate	mutant enzyme V315Y, at pH 7.4 and 37°C	Homo sapiens
0.2	-	benzyl 2-acetamido-2-deoxy-3-O-(beta-D-galactopyranosyl)-alpha-D-galactopyranoside	mutant enzyme V315Y, at pH 7.4 and 37°C	Homo sapiens
0.47	-	benzyl 2-acetamido-2-deoxy-3-O-(beta-D-galactopyranosyl)-alpha-D-galactopyranoside	mutant enzyme Y191A, at pH 7.4 and 37°C	Homo sapiens
0.62	-	CMP-N-acetyl-beta-neuraminate	mutant enzyme N170A, at pH 7.4 and 37°C	Homo sapiens
0.63	-	CMP-N-acetyl-beta-neuraminate	mutant enzyme Y191A, at pH 7.4 and 37°C	Homo sapiens
0.65	-	benzyl 2-acetamido-2-deoxy-3-O-(beta-D-galactopyranosyl)-alpha-D-galactopyranoside	mutant enzyme N170A, at pH 7.4 and 37°C	Homo sapiens
0.8	-	benzyl 2-acetamido-2-deoxy-3-O-(beta-D-galactopyranosyl)-alpha-D-galactopyranoside	mutant enzyme N147S, at pH 7.4 and 37°C	Homo sapiens
1.05	-	CMP-N-acetyl-beta-neuraminate	mutant enzyme V315A, at pH 7.4 and 37°C	Homo sapiens
1.21	-	CMP-N-acetyl-beta-neuraminate	mutant enzyme N147S, at pH 7.4 and 37°C	Homo sapiens
2.25	-	CMP-N-acetyl-beta-neuraminate	mutant enzyme C59S/C64S, at pH 7.4 and 37°C	Homo sapiens
2.28	-	benzyl 2-acetamido-2-deoxy-3-O-(beta-D-galactopyranosyl)-alpha-D-galactopyranoside	mutant enzyme C59S/C64S, at pH 7.4 and 37°C	Homo sapiens
2.32	-	benzyl 2-acetamido-2-deoxy-3-O-(beta-D-galactopyranosyl)-alpha-D-galactopyranoside	mutant enzyme S148A, at pH 7.4 and 37°C	Homo sapiens
2.42	-	benzyl 2-acetamido-2-deoxy-3-O-(beta-D-galactopyranosyl)-alpha-D-galactopyranoside	mutant enzyme V315A, at pH 7.4 and 37°C	Homo sapiens
2.5	-	CMP-N-acetyl-beta-neuraminate	mutant enzyme Y230A, at pH 7.4 and 37°C	Homo sapiens
2.7	-	CMP-N-acetyl-beta-neuraminate	mutant enzyme Y230F, at pH 7.4 and 37°C	Homo sapiens
2.97	-	benzyl 2-acetamido-2-deoxy-3-O-(beta-D-galactopyranosyl)-alpha-D-galactopyranoside	mutant enzyme Y230A, at pH 7.4 and 37°C	Homo sapiens
3.15	-	benzyl 2-acetamido-2-deoxy-3-O-(beta-D-galactopyranosyl)-alpha-D-galactopyranoside	wild type enzyme, at pH 7.4 and 37°C	Homo sapiens
3.35	-	CMP-N-acetyl-beta-neuraminate	wild type enzyme, at pH 7.4 and 37°C	Homo sapiens
3.47	-	benzyl 2-acetamido-2-deoxy-3-O-(beta-D-galactopyranosyl)-alpha-D-galactopyranoside	mutant enzyme Y230F, at pH 7.4 and 37°C	Homo sapiens
3.65	-	CMP-N-acetyl-beta-neuraminate	mutant enzyme S148A, at pH 7.4 and 37°C	Homo sapiens

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism
0.37	-	benzyl 2-acetamido-2-deoxy-3-O-(beta-D-galactopyranosyl)-alpha-D-galactopyranoside	mutant enzyme V315Y, at pH 7.4 and 37°C	Homo sapiens
0.47	-	CMP-N-acetyl-beta-neuraminate	mutant enzyme V315Y, at pH 7.4 and 37°C	Homo sapiens
0.72	-	CMP-N-acetyl-beta-neuraminate	mutant enzyme Y191A, at pH 7.4 and 37°C	Homo sapiens
1.05	-	benzyl 2-acetamido-2-deoxy-3-O-(beta-D-galactopyranosyl)-alpha-D-galactopyranoside	mutant enzyme Y191A, at pH 7.4 and 37°C	Homo sapiens
1.75	-	benzyl 2-acetamido-2-deoxy-3-O-(beta-D-galactopyranosyl)-alpha-D-galactopyranoside	mutant enzyme V315A, at pH 7.4 and 37°C	Homo sapiens
1.87	-	CMP-N-acetyl-beta-neuraminate	mutant enzyme N147S, at pH 7.4 and 37°C	Homo sapiens
3.35	-	CMP-N-acetyl-beta-neuraminate	mutant enzyme N170A, at pH 7.4 and 37°C	Homo sapiens
3.47	-	benzyl 2-acetamido-2-deoxy-3-O-(beta-D-galactopyranosyl)-alpha-D-galactopyranoside	mutant enzyme Y230A, at pH 7.4 and 37°C	Homo sapiens
3.78	-	benzyl 2-acetamido-2-deoxy-3-O-(beta-D-galactopyranosyl)-alpha-D-galactopyranoside	mutant enzyme N147S, at pH 7.4 and 37°C	Homo sapiens
5.83	-	CMP-N-acetyl-beta-neuraminate	mutant enzyme V315A, at pH 7.4 and 37°C	Homo sapiens
6.77	-	benzyl 2-acetamido-2-deoxy-3-O-(beta-D-galactopyranosyl)-alpha-D-galactopyranoside	mutant enzyme N170A, at pH 7.4 and 37°C	Homo sapiens
7.58	-	CMP-N-acetyl-beta-neuraminate	mutant enzyme S148A, at pH 7.4 and 37°C	Homo sapiens
9.85	-	benzyl 2-acetamido-2-deoxy-3-O-(beta-D-galactopyranosyl)-alpha-D-galactopyranoside	mutant enzyme Y230F, at pH 7.4 and 37°C	Homo sapiens
12.2	-	CMP-N-acetyl-beta-neuraminate	mutant enzyme Y230A, at pH 7.4 and 37°C	Homo sapiens
15.33	-	CMP-N-acetyl-beta-neuraminate	mutant enzyme Y230F, at pH 7.4 and 37°C	Homo sapiens
18.6	-	CMP-N-acetyl-beta-neuraminate	mutant enzyme C59S/C64S, at pH 7.4 and 37°C	Homo sapiens
22.07	-	benzyl 2-acetamido-2-deoxy-3-O-(beta-D-galactopyranosyl)-alpha-D-galactopyranoside	mutant enzyme S148A, at pH 7.4 and 37°C	Homo sapiens
31.6	-	CMP-N-acetyl-beta-neuraminate	wild type enzyme, at pH 7.4 and 37°C	Homo sapiens
114.17	-	benzyl 2-acetamido-2-deoxy-3-O-(beta-D-galactopyranosyl)-alpha-D-galactopyranoside	mutant enzyme C59S/C64S, at pH 7.4 and 37°C	Homo sapiens
121.15	-	benzyl 2-acetamido-2-deoxy-3-O-(beta-D-galactopyranosyl)-alpha-D-galactopyranoside	wild type enzyme, at pH 7.4 and 37°C	Homo sapiens