Kallikrein

Structure

Plasma kallikrein possesses a unique structure in vertebrates and is composed of four apple domains followed by a trypsin domain. The apple domain is a conserved protein folding with three disulfide bridges, and is often found on diverse proteins for protein-protein or protein-carbohydrate interactions. The apple domains on KLKB1 interact with the D6 domain of molecular weight kininogen (HMW-KNG), which is highly glycosylated. In teleost genomes, the KLKB1-like gene is absent and only lectins with four apple domains but no trypsin domain were found. These lectins were not found in other vertebrates, suggesting that the two genes could share the same origin but the trypsin domain was lost in the teleost lineage during evolution. Tissue kallikreins possess a single trypsin domain and are closely related to other serine proteases. It is not clear what enzyme is involved in the cleavage of KNG to form bradykinin (BK) in the fish and lamprey, given that KLKB1 and KLK are not identified.

Gene and mRNA

Plasma kallikrein (EC 3.4.21.34) is transcribed by KLKB1 located on chromosome 4q35 in humans. In mammals, KLKB1 is duplicated to form factor XI (F11) by tandem duplication.1 KLKB1 in plasma is mostly in an inactive form known as prekallikrein, and it is activated by contact activation involving HMW-KNG and factor XII. Tissue kallikreins (EC 3.4.21.35) are transcribed by the KLK genes located on chromosome 19q13 in humans. In tetrapods, the tissue kallikreins are tandemly duplicated into a large family (KLK1-KLK15 on human chromosome 19). Human KLKB1 possesses four apple domains followed by a trypsin domain and KLK possesses only a trypsin domain. The sequences of the trypsin domain are highly variable, but the catalytic domains (trypsin triads) are similar. KLK-like sequences were identified in teleosts, but the catalytic domains are mutated, indicating a possible loss of function of these enzymes and the genes became pseudogenes. Therefore, other trypsin-like enzymes may have taken the role of these KLK-like enezymes in teleosts to produce [Arg0 ]-BK from KNG.

Synthesis and release

Although KLKB1 and F11 are tandem duplicated orthologs, their regulations are different. Hepatic nuclear factor 4α (HNF4α) deletion decreased F11 but not KLKB1. Estrogen or thyroid hormone treatment increased F11 expression but not KLKB1 while a high-fat diet increased both F11 and KLKB1 expression. For KLK, the regulation of synthesis and release was most studied in detail in KLK3 and multiple androgenresponsive elements were identified upstream of KLK3.

Biological functions

KLKB1 selectively cleaves Arg/Xaa and Lys/Xaa bonds, including Lys/Arg and Arg/Ser bonds in human KNG, to release BK. It also digests plasminogen to plasmin, and participates in the surface-dependent activation of blood coagulation, fibrinolysis, and inflammation. It converts prorenin to renin to activate the RAS. Teleost KLKB1-like lectin facilitates hemagglutination by binding to the pathogen-like glycoproteins, and could be involved in immune function. KLK is highly selective to release [Lys0 ]-BK from both HMW- and LMW-KNGs, which involves the hydrolysis of Met/Xaa or Leu/Xaa. Besides acting as an enzyme for KKS, KLK is also involved in proteolytic cascades for semen liquefaction through the hydrolysis of seminogelin and the desquamation of the skin by the cleavage of cellular adhesion proteins.

Description

Plasma kallikrein possesses a unique protein structure with four apple domains and a trypsin domain, which evolved before coagulation factor XI. Tissue kallikreins are trypsinbased enzymes, and some members are highly correlated with prostate cancer. The evidence that human urine induces hypotension when injected intravenously into anesthetized dogs was first described in 1909. Two major kallikreins, plasma kallikrein (KLKB1) and tissue (glandular) kallikrein (KLK), were found in mammals, and they were transcribed by different genes. Glandular KLK was an old name and was replaced by tissue KLK in the modern nomenclature.

Chemical Properties

Kinin releasing enzyme is a substance with vasodilatory effect extracted from the submandibular gland or pancreas of pigs. It is white or off-white powder; easily soluble in water (10%), dilute alcohol, insoluble in concentrated ethanol and general organic solvents. Dissolved in water or saline into a colorless or yellowish clear solution.Ph4.5-7.8 is stable. It is unstable to heat, strong acid, strong base and oxidizer. Isoelectric point is 3.9～4.37. Generally speaking, the higher the purity, the worse the stability. The dry powder is kept at -200 for several months with no change in activity. In Ph=8 water and buffer, frozen preservation can last quite a long time without inactivation. Heavy metal ions Hg2+, Ca2+, Mn2+, Ni2+, etc. have the same degree of inhibition (Ca2+, Mg2+ has no effect on enzyme activity), mercaptan compounds and chelating agent EDTA, etc. can reverse the inhibition of metal ions on it. On the contrary, a high concentration of Ca2+ (1 molL^-{-1 can make the enzyme activity increased by 15% ~ 20%. Certain trypsin inhibitors such as peptidase, diisopropylphosphate, etc. have an inhibitory effect on pancreatic and submandibular gland kinin releasing enzyme.

Uses

Kallikrein from porcine pancreas has been used:

• as a matrix metalloproteinase-9 (MMP-9) zymogen activator
• as a component of cell culture to test its effect on rat subventricular zone (SVZ) cells and oligodendrocyte progenitor cells (OPC) proliferation and survival
• as a model enzyme to track kinetic data and visual detection limits of hydrolysis by hydrolytic enzymes in the two-phases array

General Description

Kallikrein exists as an inactive prokallikrein in the porcine pancreas. The porcine kallikrein gene region is localized on chromosome 6q12-q21.

Biochem/physiol Actions

Kallikrein active forms are generated by the enzymatic action of trypsin. It is a serine protease that mediates the activation of growth factors and substrates.

Clinical Use

Kallikreins are drug targets for the control of hypertension, inflammation, and blood coagulation diseases. They are also possible biomarkers for cancer. KLK2 and KLK3 [prostate-specific antigen (PSA)] are used as the serum biomarker for prostate cancer.