A 3d cartoon depiction of the threonine deaminase tetramer
|PDB structures||RCSB PDB PDBe PDBsum|
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Threonine ammonia-lyase, also commonly referred to as threonine deaminase or threonine dehydratase, is an enzyme responsible for catalyzing the conversion of L-threonine into alpha-ketobutyrate and ammonia. Alpha-ketobutyrate can be converted into L-isoleucine, so threonine ammonia-lyase functions as a key enzyme in BCAA synthesis.It employs a pyridoxal-5'-phosphate cofactor, similar to many enzymes involved in amino acid metabolism. It is found in bacteria, yeast, and plants, though most research to date has focused on forms of the enzyme in bacteria. This enzyme was one of the first in which negative feedback inhibition by the end product of a metabolic pathway was directly observed and studied. The enzyme serves as an excellent example of the regulatory strategies used in amino acid homeostasis.
Threonine ammonia-lyase is a tetramer of identical subunits, and is arranged as a dimer of dimers. Each subunit has two domains: a domain containing the catalytic active site and a domain with allosteric regulatory sites. The two have been shown to be distinct regions,but the regulatory site of one subunit actually interacts with the catalytic site of another subunit. Both domains contain the repeating structural motif of beta sheets surrounded by alpha helices. While the threonine binding site is not perfectly understood, structural studies do reveal how the pyridoxal phosphate cofactor is bound. The PLP cofactor is bonded to a lysine residue by means of a Schiff base, and the phosphate group of PLP is held in place by amine groups derived from a repeating sequence of glycine residues. The aromatic ring is bound to phenylalanine, and the nitrogen on the ring is hydrogen bonded to hydroxyl group-containing residues.
The mechanism of threonine ammonia-lyase is analogous to other deaminating PLP enzymes in its use of Schiff base intermediates.Initially, the amine group of threonine attacks the lysine/PLP Schiff base, displacing lysine. After deprotonation of the amino acid alpha carbon and subsequent dehydration (hence the common name threonine dehydratase), a new Schiff base is formed. This Schiff base is replaced by lysine attack, reforming the catalytically active PLP and releasing an initial alkene-containing product. This product tautomerizes, and after hydrolysis of the Schiff base, the final products are generated. After the final alpha-ketobutyrate product is generated, isoleucine is synthesized by progressing through the intermediates alpha-acetohydroxybutyrate to alpha-beta-dihydroxy-beta-methylvalerate, then to alpha-keto-beta-methylvalerate.
Threonine ammonia-lyase has been shown to not follow Michaelis-Menten kinetics, rather, it is subject to complex allosteric control.The enzyme is inhibited by isoleucine, the product of the pathway it participates in, and is activated by valine, the product of a parallel pathway. Thus, an increase in isoleucine concentration shuts off its production, and an increase in valine concentration diverts starting material (Hydroxyethyl-TPP) away from valine production. The enzyme has two binding sites for isoleucine; one has a high affinity for isoleucine and the other has a low affinity. The binding of isoleucine to the high affinity site increases the binding affinity of the low affinity site, and enzyme deactivation occurs when isoleucine binds to the low affinity site. Valine promotes enzyme activity by competitively binding to the high affinity site, preventing isoleucine from having an inhibitory effect. The combination of these two feedback methods balances the concentration of BCAAs.
Multiple forms of threonine ammonia-lyase have been observed in a variety of species of organism. In Escherichia coli , a system in which the enzyme has been studied extensively, two different forms of the enzyme are found. One is biosynthetic and resembles the enzyme characteristics presented here, while the other is degradative and functions to generate carbon fragments for energy production.The pair of isoforms has also been observed in other bacteria. In many bacteria, the biodegradative isoform of the enzyme is expressed in anaerobic conditions and is promoted by cAMP and threonine, while the biosynthetic isoform is expressed in aerobic conditions. This allows the bacterium to balance energy stores and inhibit energy-consuming synthetic pathways when energy is not abundant.
In plants, threonine ammonia-lyase is important in defense mechanisms against herbivores and is upregulated in response to abiotic stress.An adapted isoform of the enzyme with unique properties that deter herbivores is expressed in plant leaves. The catalytic domain of this isoform is extremely resistant to proteolysis, while the regulatory domain degrades readily, so upon ingestion by another organism, the threonine deamination capabilities of the enzyme go unchecked. This degrades threonine before the herbivore can absorb it, starving the herbivore of an essential amino acid. Studies of threonine ammonia-lyase in plants have also offered new strategies in the development of GMOs with increased nutritional value by increasing essential amino acid content.
Other more exotic forms of the enzyme have been found that are extremely small in size, but still retain all catalytic and regulatory functions.
There are five major fold types for PLP-dependent enzymes. Threonine ammonia-lyase is a member of the Fold Type II family, also known as the tryptophan synthase family.Though threonine ammonia-lyase does not possess substrate tunneling like tryptophan synthase does, it contains much conserved homology. Threonine ammonia-lyase is most closely related to serine dehydratase, and both possess the same general catalytic mechanism. In fact, threonine ammonia-lyase has been shown to exhibit some specificity towards serine and can convert serine into pyruvate. The regulatory domain of threonine ammonia-lyase is very similar to the regulatory domain of phosphoglycerate dehydrogenase. All of these relationships demonstrate that threonine ammonia-lyase has close evolutionary ties to these enzymes. Due to the degree of conserved structure and sequence in enzymes that recognize amino acids, it is likely that the evolutionary diversity of these enzymes came about by the matching together of individual regulatory and catalytic domains in various ways.
Threonine ammonia-lyase is not found in humans. Thus, this is one example of why humans cannot synthesize all 20 proteinogenic amino acids; in this specific case, humans cannot convert threonine into isoleucine and must consume isoleucine in the diet.The enzyme has also been studied in the past as a possible tumor suppressing agent for the previously described reasons, in that it deprives tumor cells of an essential amino acid and kills them, but this treatment has not been utilized.
Aspartate carbamoyltransferase catalyzes the first step in the pyrimidine biosynthetic pathway.
Transamination, a chemical reaction that transfers an amino group to a ketoacid to form new amino acids. This pathway is responsible for the deamination of most amino acids. This is one of the major degradation pathways which convert essential amino acids to non-essential amino acids.
Pyridoxal phosphate (PLP, pyridoxal 5'-phosphate, P5P), the active form of vitamin B6, is a coenzyme in a variety of enzymatic reactions. The Enzyme commission has catalogued more than 140 PLP-dependent activities, corresponding to ~4% of all classified activities. The versatility of PLP arises from its ability to covalently bind the substrate, and then to act as an electrophilic catalyst, thereby stabilizing different types of carbanionic reaction intermediates.
Glycogen phosphorylase is one of the phosphorylase enzymes. Glycogen phosphorylase catalyzes the rate-limiting step in glycogenolysis in animals by releasing glucose-1-phosphate from the terminal alpha-1,4-glycosidic bond. Glycogen phosphorylase is also studied as a model protein regulated by both reversible phosphorylation and allosteric effects.
A branched-chain amino acid (BCAA) is an amino acid having an aliphatic side-chain with a branch. Among the proteinogenic amino acids, there are three BCAAs: leucine, isoleucine, and valine. Non-proteinogenic BCAAs include 2-aminoisobutyric acid.
ASL is an enzyme that catalyzes the reversible breakdown of argininosuccinate (ASA) producing the amino acid arginine and dicarboxylic acid fumarate. Located in liver cytosol, ASL is the fourth enzyme of the urea cycle and involved in the biosynthesis of arginine in all species and the production of urea in ureotelic species. Mutations in ASL, resulting low activity of the enzyme, increase levels of urea in the body and result in various side effects.
Amino acid synthesis is the set of biochemical processes by which the amino acids are produced. The substrates for these processes are various compounds in the organism's diet or growth media. Not all organisms are able to synthesize all amino acids. For example, humans can only synthesize 11 of the 20 standard amino acids, and in time of accelerated growth, histidine can be considered an essential amino acid.
The acetolactate synthase (ALS) enzyme is a protein found in plants and micro-organisms. ALS catalyzes the first step in the synthesis of the branched-chain amino acids.
Serine hydroxymethyltransferase (SHMT) is a pyridoxal phosphate (PLP) (Vitamin B6) dependent enzyme (EC 126.96.36.199) which plays an important role in cellular one-carbon pathways by catalyzing the reversible, simultaneous conversions of L-serine to glycine and tetrahydrofolate (THF) to 5,10-Methylenetetrahydrofolate (5,10-CH2-THF). This reaction provides the largest part of the one-carbon units available to the cell.
Cystathionine-β-synthase, also known as CBS, is an enzyme (EC 188.8.131.52) that in humans is encoded by the CBS gene. It catalyzes the first step of the transsulfuration pathway, from homocysteine to cystathionine:
Serine dehydratase or L-serine ammonia lyase (SDH) is in the β-family of pyridoxal phosphate-dependent (PLP) enzymes. SDH is found widely in nature, but its structural and chemical properties vary greatly among species. SDH is found in yeast, bacteria, and the cytoplasm of mammalian hepatocytes. The reaction it catalyzes is the deamination of L-serine to yield pyruvate, with the release of ammonia.
Ribose-phosphate diphosphokinase is an enzyme that converts ribose 5-phosphate into phosphoribosyl pyrophosphate (PRPP). It is classified under EC 184.108.40.206.
Cystathionine gamma-lyase is an enzyme which breaks down cystathionine into cysteine, α-ketobutyrate, and ammonia. Pyridoxal phosphate is a prosthetic group of this enzyme.
In enzymology, a homoserine dehydrogenase (EC 220.127.116.11) is an enzyme that catalyzes the chemical reaction
Cystathionine beta-lyase, also commonly referred to as CBL or β-cystathionase, is an enzyme that primarily catalyzes the following α,β-elimination reaction
Methionine gamma-lyase (MGL) is an enzyme in the γ-family of PLP-dependent enzymes. It degrades sulfur-containing amino acids to α-keto acids, ammonia, and thiols. Because sulfur-containing amino acids play a role in multiple biological processes, the regulation of these amino acids is essential. Additionally, it is crucial to maintain low homocysteine levels for the proper functioning of various pathways and for preventing the toxic effects of the cysteine homologue. Methionine gamma-lyase has been found in several bacteria (Clostridiums porogenes, Pseudomonas ovalis, Pseudomonas putida, Aeromonas sp., Citrobacter intermedius, Brevibacterium linens, Citrobacter freundii, Porphyromonas gingivalis, Treponema denticola), parasitic protozoa (Trichomonas vaginalis, Entamoeba histolytica), and plants (Arabidopsis thaliana).
Acid-Induced Arginine Decarboxylase (AdiA), also commonly referred to as arginine decarboxylase, is an enzyme responsible for catalyzing the conversion of L-arginine into agmatine and carbon dioxide. The process consumes a proton in the decarboxylation and employs a pyridoxal-5'-phosphate (PLP) cofactor, similar to other enzymes involved in amino acid metabolism, such as ornithine decarboxylase and glutamine decarboxylase. It is found in bacteria and virus, though most research has so far focused on forms of the enzyme in bacteria. During the AdiA catalyzed decarboxylation of arginine, the necessary proton is consumed from the cell cytoplasm which helps to prevent the over-accumulation of protons inside the cell and serves to increase the intracellular pH. Arginine decarboxylase is part of an enzymatic system in Escherichia coli, Salmonella Typhimurium, and methane-producing bacteria Methanococcus jannaschii that makes these organisms acid resistant and allows them to survive under highly acidic medium.
In enzymology, diaminopimelate decarboxylase, also known as diaminopimelic acid decarboxylase, DAPDC, meso-diaminopimelate decarboxylase, DAP-decarboxylase, and meso-2,6-diaminoheptanedioate carboxy-lyase, is an enzyme that catalyzes the cleavage of carbon-carbon bonds in meso 2,6 diaminoheptanedioate to produce CO2 and L-lysine, the essential amino acid. It employs the cofactor pyridoxal phosphate, also known as PLP, which participates in numerous enzymatic transamination, decarboxylation and deamination reactions.
In molecular biology, the Cys/Met metabolism PLP-dependent enzyme family is a family of proteins including enzymes involved in cysteine and methionine metabolism which use PLP (pyridoxal-5'-phosphate) as a cofactor.
Glycogen phosphorylase, liver form (PYGL), also known as human liver glycogen phosphorylase (HLGP), is an enzyme that in humans is encoded by the PYGL gene on chromosome 14. This gene encodes a homodimeric protein that catalyses the cleavage of alpha-1,4-glucosidic bonds to release glucose-1-phosphate from liver glycogen stores. This protein switches from inactive phosphorylase B to active phosphorylase A by phosphorylation of serine residue 14. Activity of this enzyme is further regulated by multiple allosteric effectors and hormonal controls. Humans have three glycogen phosphorylase genes that encode distinct isozymes that are primarily expressed in liver, brain and muscle, respectively. The liver isozyme serves the glycemic demands of the body in general while the brain and muscle isozymes supply just those tissues. In glycogen storage disease type VI, also known as Hers disease, mutations in liver glycogen phosphorylase inhibit the conversion of glycogen to glucose and results in moderate hypoglycemia, mild ketosis, growth retardation and hepatomegaly. Alternative splicing results in multiple transcript variants encoding different isoforms [provided by RefSeq, Feb 2011].