Hydrolases extracellular

Hydrolases represent a significant class of therapeutic enzymes [Enzyme Commission (EC) 3.1—3.11] (14) (Table 1). Another group of enzymes with pharmacological uses has budt-ia cofactors, eg, in the form of pyridoxal phosphate, flavin nucleotides, or zinc (15). The synthases, and other multisubstrate enzymes that require high energy phosphates, are seldom available for use as dmgs because the required co-substrates are either absent from the extracellular space or are present ia prohibitively low coaceatratioas. 

The matrix metalloprotease (MMP) family of zinc hydrolases are thought to play important roles in extracellular tissue remodeling in angiogenesis and other normal physiological processes, in some inflammatory processes and in metastatic processes in cancer. Like the zinc carboxypeptidases, the MMPs also utilize a zinc-coordinated water molecule to initiate attack on the scissile amide bond of protein substrates. These enzymes are synthesized by the ribosome in a latent form composed of a catalytic domain and an N-terminal extension, referred to as the prodomain the latent, or inactive form of the enzyme is referred to as a zymogen or... 

Extracellular, exo-type poly(/3-L-malic acid) hydrolase 124... 

Brandi et al. [71] using culture fluid of Acidovorax delafieldii and cyclic 3HB oligomers were in agreement with the presence of endo-hydrolase activity of poly(3HB) depolymerases. Similar results were obtained by de Koning et al. [72] who demonstrated that covalently cross-linked poly(HAMCL) was hydrolyzed completely by P. fluorescens. It is assumed that most - if not all - extracellular poly(HA) depolymerases have endo- and exo-hydrolase activity. Depending on the depolymerase the hydrolysis products are only monomers, monomers and dimers, or a mixture of oligomers (mono- to trimers). 

Under some circumstances, lysosomal hydrolases may fail to be properly packaged in the TGN, so they enter the default pathway to the cell surface, where they are secreted. Although these hydrolases do little harm at the nearly neutral pH of most extracellular fluids, they can also be returned to lysosomes by a pathway known as receptor-mediated endocytosis. In this pathway, M6P receptors are sent to the plasma membrane, where they bind escaped lysosomal hydrolases and bring them back to lysosomes through the early and late endosomes. Receptor-mediated endocytosis is a major component of the endocytic pathways for trafficking of membrane proteins and merit more detailed consideration. 

Alice et al studied the turnover kinetics of Listeria OTonocytogenex-secreted p60 protein (a murein hydrolase) by host cell cytosolic proteasomes. J774 cells, seeded in flasks and incubated overnight in culture medium, were infected with log-phase cultures of E. monocytogenes for 30 min, washed, and incubated in culture medium for 3 h, with gentamicin (50 tg/ml) added after the first 30 min to inhibit extracellular bacterial growth. Cells then were washed and placed in methionine-free medium with spectinomycin, gentamicin, the eukaryotic protein synthesis inhibitors [cycloheximide (50 tg/mL) and anisomycin (30 tg/ml),] and 25 dVI calpain inhibitor I. After 30 min, [ S]methionine was added, and the cells were pulse-labeled for periods of 20 to 60 min. Cells... 

Sialyltransferase activity has also been described in plasma membranes from different cells, including blood platelets.285 The function of the enzyme on cell surfaces is unknown, as resialylation of membrane components directly on the cell surface is improbable, owing to the presence of CMP-Neu5Ac hydrolase in the plasma membrane, and the lack of CMP-sialic acids in the extracellular space. Furthermore, evidence has never been obtained for a role of cell-surface gly-cosyltransferases in cellular interaction according to a hypothesis of Roseman s.281... 

NG14. This mutant is capable of elaborating 15 FP units/mL with a productivity of 45 units/L/hr. The final concentration of soluble extracellular protein is 21.2 mg/mL. Samples of this enzyme preparation were sent to G. Pettersson at the University of Uppsala, Sweden, for quantification of each of the enzymes in the cellulase complex using purified antibodies to the individual enzymes. The quantitative antigen-antibody reaction showed that 600 mg/g of this enzyme preparation was one enzyme, cellobiohydrolase. This represents a yield of 13 g/L of cellobio-hydrolase, which is a 100-fold increase over the amount of cellobiohydrolase obtained with strain QM 9414 (130 mg/L) the best previously existing cellulase mutant (G. Pettersson, personal communication). 

One of the major features of solid tumors and even small deposits of tumor tissue is deficiency in the level of oxygen, because of an inadequate vascular supply. The adenosine elevation in response to hypoxia is not exclusive to tumor tissues, but, in this context, the adenosine elevation is localized to the tumor microenvironment, since the surrounding tissue is normally oxygenated. Adenosine is generated mainly by two enzymatic systems intra- or extracellularly localized 5 -nucleoti-dases and cytoplasmic S-adenosylhomocysteine hydrolase. The processes of adenosine elimination in the cell involve reactions catalyzed by adenosine deaminase and adenosine kinase (Shryock and Belardinelli 1997) yielding inosine or 5 -AMP,... 

Hydrolytic reactions are catalyzed by extracellular hydrolases and mineral surfaces (Chrost, 1990 Hoffman, 1990). For enzymatic reactions, a defined substrate or moiety must match the catalytic site of a specific enzyme. The most studied examples in aquatic systems are glycosidases, peptidases, and phosphatases (Munster and De Hann, 1998 see Chapter 13). In general, hydrolytic reactions break the relatively labile C—N and C—O bonds that link monomers, generating lower molecular weight products more suitable for microbial consumption. 

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