html PK: plasma kallikrein PPK: plasma prekallikrein TK: tissue kallikrein TproK: tissue prokallikrein HK: high molecular weight kininogen LK: low molecular weight kininogen Anti-cancer drugs  BK causes cancer cell growth by stimulating: 1. Cancer cells to grow directly 2. Matrix metalloprotease enzymes à promote cancer migration and invasion 3. Formation of new blood vessels  Many anti-cancer drugs target 1 aspect  BK antagonists target all 3

Kininogens are inhibotrs of thiol proteases. HMW kininogen plays an imp role in blood coagulation by helping to position optimally prekallikrein and factor XI next to factor XII; HMW-kininogen inhibits the thrombin- and plasmin-induced aggregation of thrombocytes; the active peptide bradykinin that is released from HMW-kininogen shows these physiological effects: influence in SM contraction, induction of hypotension, natriuresis and diuresis, decrease in blood glucose, it is a mediator of inflammation and causes increase in vascular permeability, stimulation of nociceptors, release of prostglandins (mediates inflammation) Kallikrein 1 (TK) releases Lys-bradykinin from LMW kininogen Low renal synthesis and urinary excretion of tissue kallikreins have been linked to hypertension Introduction Flow diagram: Aminopeptidase removes NH2-terminal lysine to produce bradykinin from kallidin (Lys-bradykinin) In immune and inflammatory responses, factor XIIa is activated by charged surfaces (bacterial lipopolysaccharide or damaged basement membranes). Activated factor XIIa cleaves pre-kallikrein (PK) to plasma kallikrein which acts upon HMWK to release the vasoactive nonapeptide, bradykinin. Kallikrein is serine protease Kininogens synthesized by hepatocytes, released into circulation, can be found on neutrophils, platelets, endothelial cells.

Kinins short ½ life <30s coz rapidly degraded by kininases Bradykinin (BK) has the sequence Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg, and various kininases attack different amino acid positions.(diagram) The decapeptide kallidin has the sequence Lys-Arg-Pro-Pro-Gly-Phe-Ser-ProPhe-Arg, which is equivalent to bradykinin with an extra amino acid (lysine) in the number one position Questions: -PPK made in hepatocytes à secreted into blood, circulates as complex with high mol weight kininogen. Complex is bound to external surface of neutrophil L-kininogen found on external surface of neutrophils aminopeptidases are found in plasma and surface of epithelial cells Receptors: B1 receptor normally absent but upregulated (in vitro) by bacterial products (endotoxins, lipopolysaccharides) and inflammatory mediators (TNFa) [des-Arg] kinin formed when kininase I cleaves C-terminal arginine from bradykinin and kallidin. They are G-protein coupled receptors with 7 transmembrane segments. Linking of each kinin receptor to a specific GPCR messenger system explains why bradykinin can produce diff actions on the same cell Cancer Carcinoma: malignant tumour that arises from epithelial cells Increased kinin levels have been found in these tumours increase kinin receptors associated with cell transformation by ras oncogene Tissue and plasma kallikreins hydrolyse macromol of the extracellular matrix Mesothelioma type of lung cancer caused by exposure to asbestos. When inhaled, asbestos fibers become embedded in the pleura, or outer lining of the lungs, forming scars and lesions. These lesions and scars eventually develop into pleural mesothelioma, a deadly form of lung cancer for which there is no cure. The lifespan of a person diagnosed with pleural mesothelioma is about six months to two years. Aim 1 ICC= immunocytochemistry The use of antibodies to detect proteins within cells.

Horse-radish peroxidase acts on DAB to produce an insoluble brown precipitate Gq pathway Activated Gq with GTP bound activates phospholipase C (PLC) to hydrolyze the membrane lipid PIP2, producing IP3 and diacylglycerol (DAG). IP3 diffuses to the ER, where it binds to and opens a calcium channel, releasing calcium stores from inside the ER into the cytoplasm. Calcium binds to calmodulin and calcineurin. The interaction of both DAG and calcium with PKC activates its kinase activity and the phosphorylation of many different protein targets alters their activity. The involvement of PKC in cellular proliferation and the cell cycle is indicated by the activity of tumor promoters as PKC activators. Useful info: Small cell carcinoma cells killed by dimeric kinin receptor antagonists hK1 (tissue kallikrein) induced cell migration inhibited by kallistatin (inhibitor of hK1) Effect of kallikreins on tumour cell migration and apoptosis of lung cancer cells by dimeric kinin receptor antagonists mayl have therapeutic applications Epigenetic modifications that change expression of genes involved in kallikrein-kinin cascade contribute to tumour development, invasiveness and metastasis of carcinomas of the lung