Literature summary for 5.3.1.1 extracted from

Kursula, I.; Partanen, S.; Lambeir, A.M.; Wierenga, R.K.
The importance of the conserved Arg191-Asp227 salt bridge of triosephosphate isomerase for folding, stability, and catalysis (2002), FEBS Lett., 518, 39-42.

Search for more literature for 5.3.1.1

Crystallization (Commentary)

Crystallization (Comment)	Organism
mutant enzyme R191S, hanging drop vapor diffusion method, crystal structure of the R191S mutant complexed with 2-phosphoglycolate is determined at 1.65 A resolution	Trypanosoma brucei

Protein Variants

Protein Variants	Comment	Organism
D227A	mutant enzyme with reduced stability to 3.2 M guanidinium-HCl and reduced thermal stability at 42°C. The ratio of turnover number to Km for D-glyceraldehyde 3-phosphate as substrate is 2.2fold lower than the value for the wild-type enzyme. The ratio of turnover number to Km for glycerone phosphate as substrate is 5fold lower than the value for the wild-type enzyme	Trypanosoma brucei
D227N	mutant enzyme with reduced stability to 3.2 M guanidinium-HCl and reduced thermal stability at 42°C. The ratio of turnover number to Km for D-glyceraldehyde 3-phosphate as substrate is 3.75fold lower than the value for the wild-type enzyme. The ratio of turnover number to Km for glycerone phosphate as substrate is 3.5fold lower than the value for the wild-type enzyme	Trypanosoma brucei
R191A	mutant enzyme with reduced stability to 3.2 M guanidinium-HCl and reduced thermal stability at 42°C. The ratio of turnover number to Km for D-glyceraldehyde 3-phosphate as substrate is 10.4fold lower than the value for the wild-type enzyme. The ratio of turnover number to Km for glycerone phosphate as substrate is 10.6fold lower than the value for the wild-type enzyme	Trypanosoma brucei
R191S	mutant enzyme with reduced stability to 3.2 M guanidinium-HCl and reduced thermal stability at 42°C. The ratio of turnover number to Km for D-glyceraldehyde 3-phosphate as substrate is 2fold lower than the value for the wild-type enzyme. The ratio of turnover number to Km for glycerone phosphate as substrate is 6.7fold lower than the value for the wild-type enzyme	Trypanosoma brucei

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.24	-	D-glyceraldehyde 3-phosphate	mutant enzyme S227A	Trypanosoma brucei
0.25	-	D-glyceraldehyde 3-phosphate	wild-type enzyme	Trypanosoma brucei
0.28	-	D-glyceraldehyde 3-phosphate	mutant enzyme S227N	Trypanosoma brucei
0.34	-	D-glyceraldehyde 3-phosphate	mutant enzyme R191S	Trypanosoma brucei
0.67	-	glycerone phosphate	mutant enzyme R191A	Trypanosoma brucei
0.7	-	glycerone phosphate	mutant enzyme D227N	Trypanosoma brucei
0.9	-	glycerone phosphate	mutant enzyme D227A	Trypanosoma brucei
1	-	glycerone phosphate	wild-type enzyme	Trypanosoma brucei
1.1	-	D-glyceraldehyde 3-phosphate	mutant enzyme R191A	Trypanosoma brucei
1.6	-	glycerone phosphate	mutant enzyme R191S	Trypanosoma brucei

Organic Solvent Stability

Organic Solvent	Comment	Organism
guanidine-HCl	3.2 M, half-life of wild-type enzyme: 30 min, half-life of mutant enzyme D227N: 20 min, half-life of mutant enzyme D227A: 15 min, half-life of mutant enzyme R191S: 8 min, half-life of mutant enzyme R191A: 19 min	Trypanosoma brucei

Organism

Organism	UniProt	Comment	Textmining
Trypanosoma brucei	P04789	brucei TIM	-

Renatured (Commentary)

Renatured (Comment)	Organism
protein unfolded in 3.2 M guanidinium-HCl is diluted to final concentrations of 0.32 M guanidinium-HCl, 16% recovery of wild-type enzyme after 72 h, 2.1-2.6% recovery of the mutant enzymes D227N, D227A, R191S and R191A after 72 h	Trypanosoma brucei

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
D-Glyceraldehyde 3-phosphate	-	Trypanosoma brucei	Glycerone phosphate	-	r

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
42	-	half-life of wild-type enzyme: 180 min, half-life of mutant enzyme D227N: 128 min, half-life of mutant enzyme D227A: 122 min, half-life of mutant enzyme R191S: 87 min, half-life of mutant enzyme R191A: 124 min	Trypanosoma brucei

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
63	-	glycerone phosphate	mutant enzyme R191A	Trypanosoma brucei
180	-	glycerone phosphate	mutant enzyme D227A	Trypanosoma brucei
200	-	glycerone phosphate	mutant enzyme D227N	Trypanosoma brucei
240	-	glycerone phosphate	mutant enzyme R191S	Trypanosoma brucei
1000	-	glycerone phosphate	wild-type enzyme	Trypanosoma brucei
1600	-	D-glyceraldehyde 3-phosphate	mutant enzyme S227N	Trypanosoma brucei
2500	-	D-glyceraldehyde 3-phosphate	mutant enzymeR191A	Trypanosoma brucei
2600	-	D-glyceraldehyde 3-phosphate	mutant enzyme S227A	Trypanosoma brucei
4000	-	D-glyceraldehyde 3-phosphate	mutant enzyme R191S	Trypanosoma brucei
6000	-	D-glyceraldehyde 3-phosphate	wild-type enzyme	Trypanosoma brucei

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Release 2023.1 (January 2023)