1.Activation of tyrosine hydroxylase by polyanions and salts. An electrostatic effect.
Katz IR, Yamauchi T, Kaufman S. Biochim Biophys Acta. 1976 Mar 11;429(1):84-95.
The activity of a partially purified preparation of tyrosine hydroxylase (EC 22.214.171.124) from the bovine caudate nucleus was increased by heparin, chondroitin sulfate, phosphatidylserine, polyacrylic acid, polyvinyl sulfuric acid and both poly-D-, and poly-L-glutamic acids, all polyanions. A variety of salts both activated the enzyme and prevented the activation by the polyanions. The observations that activity is increased when the enzyme interacts with salts and with macromolecules of high negative charge density are used to infer a model for these interactions and for the structural change in the enzyme that accompanies activation.
2.Influence of different cultural conditions on cellulase production by Nectria catalinensis.
Pardo AG1, Forchiassin F. Rev Argent Microbiol. 1998 Jan-Mar;30(1):20-9.
The production of the extracellular cellulolytic enzyme system (endoglucanase, exoglucanase and cellobiase) of N. catalinensis was tested with different nitrogen sources, inorganic and organic ones, in liquid culture medium with microcrystalline cellulose. The nitrogen compounds used were: potassium nitrate, sodium nitrate, ammonium nitrate, ammonium phosphate, ammonium sulphate, ammonium chloride, ammonium carbonate, ammonium acetate, ammonium tartrate, urea, casamino acids, glycine, L-alanine, L-leucine, L-proline, L-lysine, L-aspartic acid, L-glutamic acid, L-asparagine, L-glutamine, L-phenylalanine, L-tyrosine, L-tryptophan, L-methionine and L-cysteine. Among these, ammonium nitrate and ammonium tartrate gave the highest yields of cellulases in 20-day-old cultures at a concentration equivalent to 0.75 g N/l in both cases. Optimal temperature for cellulase production, growth and cellulose degradation was 23 degrees C. On the other hand, an initial pH of 6.
3.Rat osteoblasts and ROS 17/2.8 cells contain a similar protein tyrosine phosphatase.
Titus L1, Marzilli LG, Rubin J, Nanes MS, Catherwood BD. Bone Miner. 1993 Dec;23(3):267-84.
Tyrosine phosphorylation plays a central role in intracellular signaling by many hormones and growth factors. Termination of the signal is thought to involve dephosphorylation of target proteins by phosphotyrosine phosphatases (PTPase). Soluble protein PTPases from neonatal rat osteoblasts (ROBs) and rat osteosarcoma (ROS 17/2.8) cells were chromatographically distinguished and characterized using 32P-labelled glutamate/tyrosine co-polymer as substrate. Two activities from both cell types were chromatographically separable. The dominant PTPase activity in the presence of 60-125 mM salt (E1), was eluted from phosphocellulose by 180-280 mM NaCl, bound weakly to a strong anion exchange column (QAE-trisacryl), had an apparent Km for [32P]glutamate/tyrosine copolymer of 52 micrograms/ml, was enhanced (5-10-fold, ROS; 1.5-3-fold, ROB) by assay in 125 mM NaCl, had no significant alkaline, acid, or serine phosphatase activity and had an M(r) of 53,000.
4.Partial purification and characterization of protein tyrosine kinases from normal tissues.
Braun S, Abdel Ghany M, Lettieri JA, Racker E. Arch Biochem Biophys. 1986 Jun;247(2):424-32.
Three membranous protein tyrosine kinases (PTKs) have been partially purified from human placenta and pig brain. The two placental enzymes (PTK-1 and -2) are distinct with respect to solubility in detergents, molecular weight, and enzymatic properties. The brain protein tyrosine kinase resembles placental PTK-1 with respect to molecular weight and some kinetic properties. However, stimulation of brain PTK is greater with Mn2+ than with Mg2+ whereas placental PTK-1 gives higher rates with Mg2+ than with Mn2+. All three enzymes are inhibited about 50% by 0.1 M NaCl. A monoclonal antibody raised in vitro against the brain enzyme inhibits brain PTK as well as placental PTK-2, but has no effect against PTK-1 or pp60src. It thus appears that these three enzymes are distinct entities that differ from each other both kinetically and immunologically. With synthetic tyrosine-glutamic acid polymers as a substrate, protein tyrosine kinase activity can be detected in crude extracts of membranes.