Prolyl peptidases

There are several examples of peptidomic studies using iTRAQ labeling. One study identified substrates of prolyl peptidases in mouse models, and another explored the effect of diurnal variation on the endogenous peptide composition of human parotid saliva (64,65). 

For more on this family tree, see Rosenblum, J.S., Kozarich, J.W. Prolyl peptidases A serine protease subfamily with high potential for drug discovery. Curr. Opin. Chem. Biol. 2003, 7, 496-504. 

JuiUerat-Jeanneret, L., Robert, M.-C., JuiUerat, M.A., 2010. Peptides from Lactobacillus hydrolysates of bovine milk caseins inhibit prolyl-peptidases of human colon cells. Journal of Agricultural and Food Chemistry 59, 370—377. 

Other interesting examples of proteases that exhibit promiscuous behavior are proline dipeptidase from Alteromonas sp. JD6.5, whose original activity is to cleave a dipeptide bond with a prolyl residue at the carboxy terminus [121, 122] and aminopeptidase P (AMPP) from E. coli, which is a prohne-specific peptidase that catalyzes the hydrolysis of N-terminal peptide bonds containing a proline residue [123, 124]. Both enzymes exhibit phosphotriesterase activity. This means that they are capable of catalyzing the reaction that does not exist in nature. It is of particular importance, since they can hydrolyze unnatural substrates - triesters of phosphoric acid and diesters of phosphonic acids - such as organophosphorus pesticides or organophosphoms warfare agents (Scheme 5.25) [125]. 

This enzyme [EC 3.4.11.9] (also known as Xaa-Pro aminopeptidase, X-Pro aminopeptidase, proline amino-peptidase, and aminoacylproline aminopeptidase) catalyzes the hydrolysis of a peptide bond at the iV-terminus of a peptide provided that the iV-terminal amino acyl residue is linked to a prolyl residue by that peptide bond. The enzyme will also act on dipeptides and tripeptides with that same restriction. Either manganese or cobalt is needed as a cofactor. This enzyme appears to be a membrane-bound system in both mammalian and bacterial cells. The protein belongs to the peptidase family M24B. 

This zinc-dependent enzyme [EC 3.4.15.1] (also known as dipeptidyl carboxypeptidase I, dipeptidyl-dipeptidase A, kininase II, peptidase P, and carboxycathepsin) catalyzes the release of a C-terminal dipeptide at a neutral pH. The enzyme will also act on bradykinin. The presence of prolyl residues in angiotensin I and in bradykinin results in only single dipeptides being released due to the activity of this enzyme, a protein which belongs to the peptidase M2 family. The enzyme is a glycoprotein, generally membrane-bound, that is chloride ion-dependent. 

This enzyme [EC 3.4.14.1], also called cathepsin C and cathepsin J, catalyzes the hydrolysis of a peptide bond resulting in the release of an N-terminal dipeptide, XaaXbb-Xcc, except when Xaa is an arginyl or a lysyl residue, or Xbb or Xcc is a prolyl residue. This enzyme, a member of the peptidase family Cl, is a CF-dependent lysosomal cysteine-type peptidase. 

Glutamyl endopeptidase 11 [EC 3.4.21.82], also known as glutamic acid-specific protease, catalyzes the hydrolysis of peptide bonds, exhibiting a preference for Glu-Xaa bonds much more than for Asp-Xaa bonds. The enzyme has a preference for prolyl or leucyl residues at P2 and phenylalanyl at P3. Hydrolysis of Glu-Pro and Asp-Pro bonds is slow. This endopeptidase is a member of the peptidase family S2A. 

This zinc-dependent enzyme [EC 3.4.11.1], also referred to as cytosol aminopeptidase, leucyl aminopeptidase, and peptidase S, catalyzes the hydrolysis of a terminal peptide bond such that there is a release of an N-terminal amino acid, Xaa-Xbb-, in which Xaa is preferably a leucyl residue, but may be other aminoacyl residues including prolyl (although not arginyl or lysyl). Xbb may be prolyl. In addition, amino acid amides and methyl esters are also readily hydrolyzed, but the rates with arylamides are exceedingly slow. The enzyme is activated by heavy metal ions. 

This manganese-dependent enzyme [EC 3.4.11.5] catalyzes the release of an N-terminal prolyl residue from a peptide. The mammalian enzyme, which is not specific for prolyl bonds, is possibly identical with cytosol amino-peptidase [EC 3.4.11.1]. 

PROLINE REDUCTASE L-Proline to D-proline interconversion, PROLINE RACEMASE PROLIPOPROTEIN SIGNAL PEPTIDASE PROLYL 3-HYDROXYLASE PROLYL 4-HYDROXYLASE PROLYL ISOMERASE... 

The preparation of the fluoroolefin amide isosteres is reviewed. The incorporation of the amide isosteres in peptidomimetics and the influence of that isosteric substitution on biological activity on inhibition of peptidyl prolyl isomerases cyclophilin (CyP) and Pini, dipeptidyl peptidase IV/CD26 (DPP IV) and thermolysin is described. In addition, select fiuoroolefination procedures which may have utility in the construction of fluoroolefin amide isosteres are illustrated. 

The potential utility of peptides as therapeutics with clinical applications is limited by its metabolic instability or poor transmembrane mobility. Consequently, the preparation of metabolically stable peptide analogs that can either mimic or block the function of natural peptides or enzymes is an important area of medicinal chemistry research. Synthesis of fluoroolefin amide isosteres, its incorporation in peptidomimetics, and the influence of that isosteric substitution on the inhibition of several enzymes such as peptidyl prolyl isomerases, dipeptidyl peptidase IV, and thermolysin is described. Moreover, protein folding and activity... 

In 2002, Banfi and co-workers [39] further exploited this transformation to synthesize complex peptidomimetics exemplified by 29, a potent inhibitor of the serine proteases prolyl endo-peptidase [40] and Cytomegalovirus protease [41]. 

The action of these two pancreatic exopeptidases on synthetic substrates, proteins, and peptides has been reviewed in detail by Neurath (1960). The specificity requirements which were deduced from studies with synthetic peptides have been confirmed by studies with polypeptides. The structural requirements of specific substrates for both types of carboxy-peptidase are analogous except for the nature of the amino acids which contain the free, ionized a-carboxyl group at the terminus of the substrate. Carboxypeptidase B hydrolyzes most rapidly those bonds formed by terminal lysyl and arginyl residues, whereas carboxypeptidase A hydrolyzes terminal bonds formed by a variety of aromatic, neutral, or acidic amino acids. Of the natural amino acids only carboxyl-terminal prolyl residues are resistant to the action of the enzyme. The rate of hydrolysis depends upon the nature of the side chains of the amino acids which form the susceptible bonds. Thus, differences in the rate of hydrolysis of different substrates may vary several thousandfold. The methods for application of these peptidases to hydrolysis of proteins have been discussed in detail by Canfield and Anfinsen (1963). 

Peptidases such as aminopeptidases and diamino-peptidases are at lower concentrations in the colon than in the small intestine. However, it has been shown that prolyl endoprotease and collagenase activities are five to six times higher in the colon than in the small intestine. TRH is more readily hydrolyzed to deaminated TRH in colonic homogenates compared with small intestine and rectum. Therefore, peptide drugs that are substrates for prolyl endoprotease and collagenase will likely be degraded in the colon and may not be suitable for colon delivery. 

Clan SC contains peptidases with the a/P hydrolase fold bearing the catalytic triad in the order Ser, Asp, His. This clan includes the families (characteristic member in parentheses) S9 (prolyl oligopeptidase), S10 (carboxypeptidase C), S15 (Xaa-Pro dipeptidyl -peptidase), S28 (lysosomal Pro-Xaa carboxypeptidase), S33 (prolyl amino-peptidase), and S37 (Streptomyces PS-10 peptidase). The characteristic catalytic dyad Ser, Lys of dan SE is represented by the motif Ser-Xaa-Xbb-Lys, and the fold consists of helices and an a + P sandwich. The families of this clan Sll (penicillin-binding protein 5), S12 (Streptomyces R61 D-Ala-D-Ala carboxypeptidase), S13 (penicillinbinding protein 4) are involved in the biosynthesis, turnover and lysis of bacterial cell walls. 

Goossens, R VanHoof, G. De Meester, I. Augustyns, K. Borloo, M. Tourwe, D. Haemers, A. Scharpe, S. Development and evaluation of peptide-based prolyl oligo-peptidase inhibitors. Introduction of A-benzyloxycarbonyl-prolyl-3-fluoropyrrolidine as a lead in inhibitor design. Eur. J. Biochem. 1997, 250, 177-183. 

Signal peptidase Prolyl 4-hydroxylase Prolyl 3-hydroxylase Lysyl hydroxylase Hydroxylysyl galactosyl-transferase Hydroxylysyl glucosyMransferase Oligosaccharyl transferase Protein disulphide isomerase Prolyl-peptidyl cis/trans isomerase BiP... 

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