Literature summary for 2.3.2.13 extracted from

Ha, H.J.; Kwon, S.; Jeong, E.M.; Kim, C.M.; Lee, K.B.; Kim, I.G.; Park, H.H.
Structure of natural variant transglutaminase 2 reveals molecular basis of gaining stability and higher activity (2018), PLoS ONE, 13, e0204707 .

Cloned(Commentary)

Cloned (Comment)	Organism
recombinant expression of His-tagged full-length wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)	Homo sapiens

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant His-tagged enzyme, hanging drop vapor diffusion method, mixing of 0.001 ml of 10 mg/ml protein in 20 mM Tris, pH 8.0, and 150 mM NaCl with 0.001 ml of reservoir solution containing 20 mM MES, pH 6.2, 150 mM sodium chloride, 5mM MgCl2, 4% PEG 3350, and 5 mM DTT, and 20% glycerol, equilibration against 0.4 ml of the reservoir solution, 20°C, X-ray diffraction structrue determination and analysis at 3.2 A resolution	Homo sapiens

Crystallization (Comment)

Organism

purified recombinant His-tagged enzyme, hanging drop vapor diffusion method, mixing of 0.001 ml of 10 mg/ml protein in 20 mM Tris, pH 8.0, and 150 mM NaCl with 0.001 ml of reservoir solution containing 20 mM MES, pH 6.2, 150 mM sodium chloride, 5mM MgCl2, 4% PEG 3350, and 5 mM DTT, and 20% glycerol, equilibration against 0.4 ml of the reservoir solution, 20°C, X-ray diffraction structrue determination and analysis at 3.2 A resolution

Homo sapiens

Protein Variants

Protein Variants	Comment	Organism
G224V	a putative natural mutant variant of isozyme TG2, mutant TG2 G224V gains higher stability and Ca2+-dependent activity compared to the G224 form. Possibly the G224V form, rather than the G224 form, is the natural TG2 variant	Homo sapiens
additional information	comparison of wild-type with G224V mutant enzyme structure and actives sites, overview	Homo sapiens

Inhibitors

Inhibitors	Comment	Organism	Structure
EDTA	complete inhibition at 10 mM	Homo sapiens
GTP	the protein transamidase activity of TG2 is positively regulated by calcium and negatively regulated by GTP	Homo sapiens

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Ca2+	dependent on, the protein transamidase activity of TG2 is positively regulated by calcium and negatively regulated by GTP. The wild-type and mutant enzymes both contain six Ca2+ binding sites	Homo sapiens

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
80240	-	recombinant His-tagged enzyme mutant G224V with bound GTP, gel filtration	Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
protein glutamine + alkylamine	Homo sapiens	-	protein N5-alkylglutamine + NH3	-	?

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	P21980	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged full-length wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gelfiltration, followed by anion exchange chromatography, a second step of gel filtration, and ultrafiltration	Homo sapiens

Source Tissue

Source Tissue	Comment	Organism	Textmining

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
N,N'-dimethyl casein glutamine + biotinylated pentylamine	the enzymatic activity is determined by measuring the fluorescence of biotinylated pentylamine crosslinking to N,N'-dimethyl casein upon exposure	Homo sapiens	?	-	?
protein glutamine + alkylamine	-	Homo sapiens	protein N5-alkylglutamine + NH3	-	?

Subunits

Subunits	Comment	Organism
monomer	1 * 80240, recombinant His-tagged enzyme mutant G224V with bound GTP, SDS-PAGE	Homo sapiens

Synonyms

Synonyms	Comment	Organism
TG2	-	Homo sapiens
transglutaminase 2	-	Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
37	-	assay at	Homo sapiens

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.5	-	assay at	Homo sapiens

General Information

General Information	Comment	Organism
malfunction	failure in the regulation of TG2 activities is associated with many human diseases, including inflammatory disease, celiac disease, neurodegenerative disease, diabetes, tissue fibrosis, and cancers. Comparison of wild-type with G224V mutant enzyme structrue and actives sites, overview. Since the active site cysteine (C277)-containing alpha-helix, whose location may be important for TG2 activity, is interacted with a neighboring intramolecular alpha-helix, which contains residue 224, replacement of G with V will have an effect on the transamidase activity of TG2. Decreased activity of G224 may be due to the higher chance of location shift on active site cysteine (C277) because of the loss of the hydrophobic cluster anchored by V224	Homo sapiens
physiological function	transglutaminase 2 (TG2) is a multi-functional protein that possesses various biological activities, including protein cross-linking activity, GTPase activity, protein disulfide isomerase activity, kinase activity, and scaffold activity. Because of its various functions, TG2 is involved in many important cellular processes, including apoptosis, angiogenesis, wound healing, neuronal regeneration, and bone development. Isozyme TG2 function differs according to its location in the cell. In the cytosol, TG2 acts as a signal transfer molecule that transmits a receptor signal to an intracellular effector through GTP hydrolysis. When it is secreted into the extracellular environment, TG2 functions as a cross-linking enzyme in the matrix. This protein transamidase activity of TG2 is positively regulated by calcium and negatively regulated by GTP	Homo sapiens