Tissue transglutaminase

TGM2
Available structures PDB Ortholog search: PDBe RCSB List of PDB id codes 4PYG, 1KV3, 2Q3Z, 3LY6, 3S3J, 3S3P, 3S3S
Identifiers
Aliases	TGM2, G-ALPHA-h, GNAH, HEL-S-45, TG2, TGC, TG(C), transglutaminase 2
External IDs	OMIM: 190196 MGI: 98731 HomoloGene: 3391 GeneCards: TGM2
Gene ontology Molecular function • transferase activity • transferase activity, transferring acyl groups • protein-glutamine gamma-glutamyltransferase activity • protein binding • metal ion binding • GTP binding • protein domain specific binding Cellular component • cytoplasm • intrinsic component of plasma membrane • cytosol • extracellular exosome • focal adhesion • mitochondrion • endoplasmic reticulum • plasma membrane • extracellular matrix Biological process • apoptotic cell clearance • phospholipase C-activating G-protein coupled receptor signaling pathway • salivary gland cavitation • negative regulation of endoplasmic reticulum calcium ion concentration • positive regulation of mitochondrial calcium ion concentration • peptide cross-linking • positive regulation of cell adhesion • branching involved in salivary gland morphogenesis • positive regulation of inflammatory response • positive regulation of I-kappaB kinase/NF-kappaB signaling • positive regulation of cytosolic calcium ion concentration involved in phospholipase C-activating G-protein coupled signaling pathway • isopeptide cross-linking via N6-(L-isoglutamyl)-L-lysine • positive regulation of apoptotic process • protein homooligomerization • positive regulation of smooth muscle cell proliferation • blood vessel remodeling Sources:Amigo / QuickGO
RNA expression pattern
More reference expression data
Orthologs
Species	Human	Mouse
Entrez	7052	21817
Ensembl	ENSG00000198959	ENSMUSG00000037820
UniProt	P21980	P21981
RefSeq (mRNA)	NM_004613 NM_198951 NM_001323316 NM_001323317 NM_001323318	NM_009373
RefSeq (protein)	NP_004604.2 NP_945189.1 NP_001310245.1	NP_033399.1
Location (UCSC)	Chr 20: 38.13 – 38.17 Mb	Chr 2: 158.12 – 158.15 Mb
PubMed search	[1]	[2]
Wikidata

Tissue transglutaminase (abbreviated as tTG or TG2) is a 78-kDa, calcium-dependent enzyme (EC 2.3.2.13) of the protein-glutamine γ-glutamyltransferases family (or simply transglutaminase family).^[3][4] Like other transglutaminases, it crosslinks proteins between an ε-amino group of a lysine residue and a γ-carboxamide group of glutamine residue, creating an inter- or intramolecular bond that is highly resistant to proteolysis (protein degradation). Aside from its crosslinking function, tTG catalyzes other types of reactions including deamidation, GTP-binding/hydrolyzing, and isopeptidase activities.^[5] Unlike other members of the transglutaminase family, tTG can be found both in the intracellular and the extracellular spaces of various types of tissues and is found in many different organs including the heart, the liver, and the small intestine. Intracellular tTG is abundant in the cytosol but smaller amounts can also be found in the nucleus and the mitochondria.^[4] Intracellular tTG is thought to play an important role in apoptosis.^[6] In the extracellular space, tTG binds to proteins of the extracellular matrix (ECM),^[7] binding particularly tightly to fibronectin.^[8] Extracellular tTG has been linked to cell adhesion, ECM stabilization, wound healing, receptor signaling, cellular proliferation, and cellular motility.^[4]

tTG is particularly notable for being the autoantigen in celiac disease, a lifelong illness in which the consumption of dietary gluten causes a pathological immune response resulting in the inflammation of the small intestine and subsequent villous atrophy.^[9][10][11]

Structure

Gene

The human tTG gene is located on the 20th chromosome (20q11.2-q12).

Protein

TG2 is a multifunctional enzyme that belongs to transglutaminases which catalyze the crosslinking of proteins by epsilon-(gamma-glutamyl)lysine isopeptide bonds.^[12] Crystal structures of TG2 with bound GDP, GTP, or ATP have demonstrated that these forms of TG2 adopt a "closed" conformation, whereas TG2 with the active site occupied by an inhibitory gluten peptide mimic or other similar inhibitors adopts an "open" conformation. In the open conformation the four domains of TG2 are arranged in an extended configuration, whereas in the closed conformation the two C-terminal domains are folded in on the catalytic core domain. The N-terminal domain only shows minor structural changes between the two different conformations.^[13]

Mechanism

The catalytic mechanism for crosslinking in human tTG involves the thiol group from a Cys residue in the active site of tTG.^[4] The thiol group attacks the carboxamide of a glutamine residue on the surface of a protein or peptide substrate, releasing ammonia, and producing a thioester intermediate. The thioester intermediate can then be attacked by the surface amine of a second substrate (typically from a lysine residue). The end product of the reaction is a stable isopeptide bond between the two substrates (i.e. crosslinking). Alternatively, the thioester intermediate can be hydrolyzed, resulting in the net conversion of the glutamine residue to glutamic acid (i.e. deamidation).^[4] The deamidation of glutamine residues catalyzed by tTG is thought to be linked to the pathological immune response to gluten in celiac disease.^[10] A schematic for the crosslinking and the deamidation reactions is provided in Figure 1.

Figure 1: Transamidation (crosslinking) and deamidation mechanisms of tissue transglutaminase

Regulation

Crosslinking activity by tTG requires the binding of Ca²⁺ ions.^[14] Multiple Ca²⁺ can bind to a single tTG molecule.^[4] In contrast, the binding of one molecule of GTP or GDP inhibits the crosslinking activity of the enzyme.^[14] Therefore, intracellular tTG is mostly inactive due to the relatively high concentration of GTP/GDP and the low levels of calcium inside the cell.^[4][10] Although extracellular tTG is expected to be active due to the low concentration of guanine nucleotides and the high levels of calcium in the extracellular space, evidence has shown that extracellular tTG is mostly inactive.^[4][10][14] Recent studies suggest that extracellular tTG is kept inactive by the formation of a disulfide bond between two vicinal Cys residues. Therefore, oxidation/reduction of the disulfide bond serves as a third allosteric regulatory mechanism (along with GTP/GDP and Ca²⁺) for the activation of tTG.^[10] Thioredoxin has been shown to activate extracellular tTG by reducing the disulfide bond.^[14] Recent studies have suggested that interferon-γ may serve as an activator of extracellular tTG in the small intestine; these studies have a direct implication to the pathogenesis of celiac disease.^[10] Activation of tTG has been shown to be accompanied by large conformational changes, switching from a compact (inactive) to an extended (active) conformation. (see Figure 2)^[14][15][16]

Figure 2: Compact (inactive) and extended (active) conformations of tTG

Function

tTG is expressed ubiquitously. It requires calcium as a cofactor for transamidation activity. Transcription is increased by retinoic acid. Among its many supposed functions, it appears to play a role in wound healing, apoptosis, and extracellular matrix development^[9]

tTG is thought to be involved in the regulation of the cytoskeleton by crosslinking various cytoskeletal proteins including myosin, actin, and spectrin.^[17] Evidence shows that intracellular tTG crosslinks itself to myosin. It is also believed that tTG may stabilize the structure of the dying cells during apoptosis by polymerizing the components of the cytoskeleton, therefore preventing the leakage of the cellular contents into the extracellular space.^[5]

tTG also has GTPase activity:^[3] In the presence of GTP, it suggested to function as a G protein participating in signaling processes.^[18] Besides its transglutaminase activity, tTG is proposed to also act as kinase,^[19] and protein disulfide isomerase,^[20] and deamidase.^[21] This latter activity is important in the deamidation of gliadin peptides, thus playing important role in the pathology of coeliac disease.

Clinical significance

tTG is best known for its link with celiac disease.^[11] Anti-transglutaminase antibodies result in a form of gluten sensitivity in which a cellular response to Triticeae glutens that are crosslinked to tTG are able to stimulate transglutaminase specific B-cell responses that eventually result in the production of anti-transglutaminase antibodies IgA and IgG.^[22]

tTG is believed to be involved in several neurodegenerative disorders including Alzheimer, Parkinson and Huntington diseases.^[23][24] Such neurological diseases are characterized in part by the abnormal aggregation of proteins due to the increased activity of protein crosslinking in the affected brain.^[25] Additionally, specific proteins associated with these disorders have been found to be in vivo and in vitro substrates of tTG.^[5] Although tTG is up regulated in the areas of the brain affected by Huntington's disease, a recent study showed that increasing levels of tTG do not affect the onset and/or progression of the disease in mice.^[26]

Recent studies suggest that tTG also plays a role in inflammation and tumor biology.^[9] tTG expression is elevated in multiple cancer cell types and is implicated in drug resistance and metastasis due to its ability to promote mesenchymal transition and stem cell like properties.

Diagnostic

Serology for anti-tTG antibodies has superseded older serological tests (anti-endomysium, anti-gliadin, and anti-reticulin) and has a strong sensitivity (99%) and specificity (>90%) for identifying celiac disease. Modern anti-tTG assays rely on a human recombinant protein as an antigen.^[27]

Therapeutic

It's still experimental to use tTG as a form of surgical glue. It is also being studied as an attenuator of metastasis in certain tumors.^[9]

Interactions

TG2 participates in both enzymatic and non-enzymatic interactions. Enzymatic interactions are formed between TG2 and its substrate proteins containing the glutamine donor and lysine donor groups in the presence of calcium. Substrates of TG2 are known to affect TG2 activity, which enables it to subsequently execute diverse biological functions in the cell. However, the importance of non-enzymatic interactions in regulating TG2 activities is yet to be revealed. Recent studies indicate that non-enzymatic interactions play physiological roles and enable diverse TG2 functions in a context-specific manner.^[28]

Mouse Mutant Alleles for Tgm2
Marker Symbol for Mouse Gene. This symbol is assigned to the genomic locus by the MGI	Tgm2
Mutant Mouse Embryonic Stem Cell Clones. These are the known targeted mutations for this gene in a mouse.	Tgm2tm1a(KOMP)Wtsi
Example structure of targeted conditional mutant allele for this gene
These Mutant ES Cells can be studied directly or used to generate mice with this gene knocked out. Study of these mice can shed light on the function of Tgm2: see Knockout mouse

References

External links

• Endomysial antibodies
• A collection of substrates and interaction partners of TG2 is accessible in the TRANSDAB, an interactive transglutaminase substrate database.

PDB gallery
1kv3: HUMAN TISSUE TRANSGLUTAMINASE IN GDP BOUND FORM