Calcium-dependent hydrolases

These three catalytic functionalities are similar in practically all hydrolytic enzymes, but the actual functional groups performing the reactions differ among hydrolases. Based on the structures of their catalytic sites, hydrolases can be divided into five classes, namely serine hydrolases, threonine hydrolases, cysteine hydrolases, aspartic hydrolases, and metallohydrolases, to which the similarly acting calcium-dependent hydrolases can be added. Hydrolases of yet unknown catalytic mechanism also exist. 

The aspartic hydrolases, metallohydrolases, and calcium-dependent hydrolases, which activate (i.e., render more nucleophilic) a H20 molecule and allow it to attack the substrate. Here, no covalent complex is formed with the enzyme. 

Thus, calcium-dependent hydrolases show clear structural and catalytic similarities with metallohydrolases, both in the role played by a divalent cation and in the mechanism of H20 activation. 

In the other subdivision, water activation occurs in the first step of the enzymatic cycle. This activation is achieved by a carboxylate group in aspartic hydrolases (Fig. 3.10), Zn2+ and a carboxy group in metallopep-tidases (Fig. 3.12 ), a histidine side chain in calcium-dependent hydrolases (Fig. 3.14), or a Zn2+ in carbonic anhydrase (Fig. 3.15). The substrate, on the other hand, is polarized (activated) by a carboxy group in aspartic hydrolases or by a cation in metallopeptidases and calcium-dependent hydrolases. In this manner, the reactivity of both the water molecule and the substrate is enhanced and fine-tuned to drive formation of a tetrahedral intermediate that will break down to form the hydrolysis products. 

Jakubowski, H. (2000). Calcium-dependent human serum homocysteine thiolactone hydrolase. A protective mechanism against protein N-homocysteinylation. J. Biol. Chem. 275 3957-62. 

A few enzymes, such as the previously mentioned CNP, are believed to be fairly specific for myelin/oligodendro-cytes. There is much more in the CNS than in peripheral nerve, suggesting some function more specialized to the CNS. In addition, a unique pH 7.2 cholesterol ester hydrolase is also enriched in myelin. On the other hand, there are many enzymes that are not myelin-specific but appear to be intrinsic to myelin and not contaminants. These include cAMP-stimulated kinase, calcium/calmodulin-dependent kinase, protein kinase C, a neutral protease activity and phosphoprotein phosphatases. The protein kinase C and phosphatase activities are presumed to be responsible for the rapid turnover of MBP phosphate groups, and the PLP acylation enzyme activity is also intrinsic to myelin. 

The catalytic activity of porcine pancreatic a-amylase, another member of the glucosyl hydrolase family, is metal cofactor-dependent [53]. A calcium ion-binding site is located at the interface of an antiparallel f-sheet, inserted in one of the loop regions of the /ia-barrel, and the core structure. In the calcium-bound state, the insertion stabilizes the substrate-binding site, and indirectly constrains part of the active site in a catalytically competent conformation. 

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