Prolyl peptidases: a serine protease subfamily with high potential for drug discovery

Rosenblum, Jonathan S.; Kozarich, John W.

doi:10.1016/s1367-5931(03)00084-x

Cited by 278 publications

(208 citation statements)

References 63 publications

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“…Therefore, for the cleavage of Pro-Xaa peptide bonds (where Xaa is any amino acid) a specialised group of proteases, the prolyl peptidases, have evolved and their activity in vivo may have important physiological significance (Rosenblum & Kozarich, 2003). In this report we have described the purification, sequencing and properties of a novel prolyl peptidase from bovine serum.…”

Section: Discussionmentioning

confidence: 99%

“…The prolyl peptidases are a unique class of proteases, gaining much attention of late, that possess the ability to cleave proteins and peptides after proline residues. This group includes dipeptidyl peptidase IV, seprase/fibroblast activation protein α, DPP7, DPP8, DPP9, prolyl carboxypeptidase while more distant members include prolyl oligopeptidase (Rosenblum & Kozarich, 2003). Seprase or fibroblast activation protein (FAP) is an integral membrane serine peptidase, which has been shown to have gelatinase activity.…”

Section: Introductionmentioning

confidence: 99%

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Purification, identification and characterisation of seprase from bovine serum

Collins

McMahon

O’Brien

et al. 2004

The International Journal of Biochemistry & Cell Biology

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ABBREVIATIONSAMC, 7-amino-4-methylcoumarin; BCA, bicinchoninic acid; CPC, calcium phosphate cellulose; DFP, diisopropylfluorophosphate; FPLC, fast protein liquid chromatography; HIC, hydrophobic interaction chromatography; PO, prolyl oligopeptidase; TFA, trifluoroacetic acid; Z, N-benzyloxycarbonyl; ZIP, Z-Pro-prolinal-insensitive Z-Gly-Pro-AMC-hydrolysing peptidase. ABSTRACTThe study and identification for the first time of a soluble form of a Seprase activity from bovine serum is presented. To date, this activity has only been reported to be an integral membrane protease but has been known to shed from its membrane. The activity was purified 30,197-fold to homogeneity, using a combination of column chromatographies, from bovine serum. Inhibition by DFP, resulting in an IC 50 of 100nM confirms classification as a serine protease. The protease after separation and visualisation by native PAGE was subjected to tryptic digestion and the subsequent peptides sequenced. Each peptide sequenced was found to be present in the primary structure of Seprase/Fibroblast Activation Protein (FAP), a serine gelatinase specific for proline-containing peptides and macromolecules. Substrate specificity studies using kinetic, RP-HPLC and LC-MS analysis of synthetic peptides suggest that this peptidase has an extended substrate-binding region in addition to the primary specificity site S 1 . This analysis revealed at least five subsites to be involved in enzyme-substrate binding, 2 with the smallest peptide cleaved being a tetrapeptide. A proline residue in position P 1 was absolutely necessary therefore showing high primary substrate specificity for the Pro-X bond, while a preference for a hydrophobic residue at the C-terminal end of the scissile bond (P 1 ʹ′) was evident. The enzyme also showed complete insensitivity to the prolyl oligopeptidase specific inhibitors, JTP-4819, Fmoc-Ala-pyrrCN and Z-Phe-Pro-BT. To date, no physiological substrate has clearly been defined for this protease but its ability to effectively degrade gelatin suggests a candidate protein substrate in vivo and a possible role in extracellular matrix protein degradation.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Purification, identification and characterisation of seprase from bovine serum

Collins

McMahon

O’Brien

et al. 2004

The International Journal of Biochemistry & Cell Biology

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“…Members of the S9B/DPPIV family are serine amino peptidases with the unique ability to cleave off N-terminal dipeptides from proteins/peptides having a proline residue at position 2 (X aa P). Four active members of this family are known so far: the two membrane-bound cell-surface members dipeptidyl peptidase IV (DPPIV) and the Fibroblast activation protein alpha (FaP), as well as the two soluble members DPP8 and 1 3 DPP9 (reviewed in [3][4][5][6]). Dimerization is a common feature for members of the S9B/DPPIV family [6][7][8][9][10][11] and is essential for enzymatic activity [12][13][14][15][16].…”

Section: Introductionmentioning

confidence: 99%

The amino terminus extension in the long dipeptidyl peptidase 9 isoform contains a nuclear localization signal targeting the active peptidase to the nucleus

Justa-Schuch

Möller

Geiss‐Friedlander

2014

Cell. Mol. Life Sci.

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“…In recent years, much attention has been given in the medical research field to the development of inhibitors for this type of enzyme, in particular for DPP IV (Rosenblum and Kozarich, 2003). Such inhibitors, when available, could easily be tested in the coagulation assays for their inhibitory activity of the H. contortus PCPs.…”

Section: Discussionmentioning

confidence: 99%

The intestinal contortin structure in Haemonchus contortus: An immobilised anticoagulant?

Geldhof

Knox

2008

International Journal for Parasitology

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Contortin was the first intestinal antigen of the sheep parasite Haemonchus contortus which induced significant levels of protection when used to vaccinate lambs. This antigen is present in the intestine of L4 and adult worms as a helical polymeric structure attached to the luminal surface of the intestinal cells. However, the nature of the protein itself and its function have never been reported. In the present study, contortin was isolated and analysed by peptide mass fingerprint and LC/MS-MS. These analyses indicated that contortin comprises two major proteins, Hc-PCP1 and Hc-PCP2, with homology to prolyl-carboxypeptidases. The two proteins show 64% amino acid sequence identity to each other and both are comprised of two prolyl-carboxypeptidase S28 type domains organised in a tandem repeat. The transcripts of both genes are present from the L4 stage onwards, coinciding with the onset of blood-feeding. Addition of contortin to a fibrinogen solution significantly inhibited blood coagulation in a dose-dependent manner. Mass-spectrometry indicated that the contortin-enriched fraction degraded the C-terminal end of the fibrinogen alpha-chain, which was shown previously to be essential for clot formation. The process happens within seconds after addition and can be inhibited by the dipeptidyl-peptidase IV inhibitors Diprotin A and Bt-PEG-Glu-Pro P (OPh) 2 . These data suggest that the prolyl-carboxypeptidases are intestinal anticoagulants used by H. contortus to interfere with blood coagulation. Ó

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Prolyl peptidases: a serine protease subfamily with high potential for drug discovery

Cited by 278 publications

References 63 publications

Purification, identification and characterisation of seprase from bovine serum

Purification, identification and characterisation of seprase from bovine serum

The amino terminus extension in the long dipeptidyl peptidase 9 isoform contains a nuclear localization signal targeting the active peptidase to the nucleus

The intestinal contortin structure in Haemonchus contortus: An immobilised anticoagulant?

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