Published in BBA - Enzymology
Pro-Gly-ArgCH 2Cl, a reagent corresponding to the C-terminal sequence generated in plasminogen on activation by urokinase (EC 3.4.99.26) and probably by other plasminogen activators, was prepared. Pro-Gly-ArgCH 2Cl was effective in the inactivation of urokinase at the 10 −6 M level ( K i 68 μM and λ 2 0.47 min −1). In contrast, only a slow inactiva...
Published in BBA - Protein Structure
The low field portion of the 360 MHz 1H nuclear magnetic resonance spectrum of phenylmethanesulfonyl-subtilisin Novo (EC 3.4.21.14) has been studied as a function of pH. Analysis of the pH-induced chemical shift changes occurring between 6 to 7 ppm revealed five classes of ionizable residues with p K values (uncorrected) of 10.3, 10.7, 10.7, 10.8, ...
Published in BBA - Enzymology
Prekallikrein was purified from guinea-pig plasma. The prekallikrein appeared homogeneous as a single-chain protein on polyacrylamide gels in the presence of sodium dodecyl sulfate (SDS) and β-mercaptoethanol. The apparent molecular weight was 52000 by SDS-polyacrylamide gel electrophoresis, 99 000 by gel filtration on a Sephadex G-150 column and 8...
Published in BBA - Enzymology
Dipeptidyl peptidase II (dipeptidylpeptide hydrolase, EC 3.4.14.2), previously known as dipeptidyl aminopeptidase II, was shown to be present in relatively high concentrations in bovine dental pulp. Based on the DNA content of tissue homogenates, the fibroblasts of this connective tissue appeared to contain more dipeptidyl peptidase II than the cel...
Published in BBA - Protein Structure
‘Thermitase’ (EC 3.4.21.14), a thermostable extracellular serine protease from Thermoactinomyces vulgaris, binds one calcium ion with a dissociation constant of about 10 −4 M at 25°C and pH 7.5 to 3.5. In addition, two calcium ions are bound more tightly to the enzyme, as shown by experiments with a calcium-selective electrode. The single most weak...
Genes and Development / 9 / 2 / 139-154 / GEDEE
In the avian model of myopia, retinal image degradation quickly leads to ocular enlargement. We now give evidence that regionally specific changes in ocular size are correlated with both biomechanical indices of scleral remodeling, e.g. hydration capacity and with biochemical changes in proteinase activities. The latter include a 72 kDa matrix meta...
Published in Development genes and evolution
We have studied gene expression during ascidian embryonic development using the technique of differential display and isolated partial cDNA sequences of 12 genes. Developmental regulation of these genes has been confirmed by northern hybridization analysis. Further cDNA cloning and sequence analysis of an mRNA that is present during gastrulation, n...
Published in Immunogenetics
We cloned and sequenced a mouse gene encoding a new type of membrane bound serine protease (epithin) containing a multidomain structure. The initial cDNA clone was found previously in a polymerase chain reaction (PCR)-based subtractive library generated from fetal thymic stromal cells, and the message was shown to be highly expressed in a thymic ep...
Published in Extremophiles
A protease of a molecular mass of approximately 30 kDa was isolated and purified from the haloalkaliphilic archaeon Natronomonas (formerly Natronobacterium) pharaonis. The enzyme hydrolyzed synthetic peptides, preferentially at the carboxyl terminus of phenylalanine or leucine, as well as large proteins. Hydrolysis occurred over the range of pH fro...
