Abstract
The discovery of mammalian subtilases, proprotein convertases (PCs) or subtilisin-like proprotein convertases (SPCs), in 1990 was a result of sustained efforts in searching for enzyme/s responsible for maturation of inactive protein precursors. Since then, seven PCs have so far been discovered that cleave at the carboxy-terminal of a basic amino acid characterized by the consensus sequence Arg/Lys/His-X-X/Lys/Arg-Arg↓, where X denotes any amino acid other than Cys. Two additional PC subtypes—called subtilisin kexin isozyme 1 (SKI-1) or site 1 protease (S1P) and neural apoptosis regulated convertase 1 (NARC-1), also known as PCSK9—that cleave at the carboxy terminus of nonbasic amino acids were discovered later. Numerous studies revealed various important functional roles of PCs in health and diseases such as tumorigenesis, diabetes, viral infections, bacterial pathogenesis, atherosclerosis, and neurodegenarative diseases such as Alzheimer’s. Owing to these findings, PCs became a promising frontier for treatment of diverse pathologies. Thus modulation of PC activity with designed inhibitors is an attractive proposition not only for intervention of diseases, but also for biochemical characterization of these enzymes. Various physiological and bioengineered proteins as well as small molecules such as peptide, peptidomimetic, and nonpeptide compounds as inhibitors of PCs have been described in the literature. Among the strategies used for design of PC inhibitors, the most successful is the one based on bioengineered serpin proteins, of which the best example is α1-PDX, the double mutant variant of α1-antitrypsin (from A355IPM358 to R355IPR358). Others include small peptide inhibitors with C-terminal carboxyl function modified with a potent neucleophile or those containing pseudo or isosteric peptide bond at the scissile site of a suitable peptide substrate. Among nonpeptide PC inhibitors, the number is very limited. So far, these include 20-carbon atoms containing alicyclic diterpenes of andrographolide family and heterocyclic compounds that are ligands of Zn2+ and Cu2+ ions. Overall, these molecules display only a modest enzyme inhibition; however, they may serve as important lead structures for further development of more potent and specific nonpeptide PC inhibitors as potential therapeutic agents. Many PC inhibitors display their functional properties in proliferation, fertilization, tumorigenesis, obesity, embryogenesis, or diabetes via their inhibitory action on PC activities.
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References
Seidah NG, Chrétien M (1999) Proprotein and prohormone convertases: a family of subtilases generating diverse bioactive polypeptides. Brain Res 848:45–62
Thomas G (2002) Furin at the cutting edge: from protein traffic to embryogenesis and disease. Nat Rev 3:753–766
Steiner DF (1998) The proprotein convertases. Curr Opin Chem Biol 2:31–39
Chrétien M, Li CH (1967) Isolation, purification and characterization of g-lipotropic hormone from sheep pituitary glands. Can J Biochem 45:1163–1174
Steiner DF, Cunningham D, Spiegelman L, Aten B (1967) Insulin biosynthesis: evidence for a precursor. Science 157:697–699
Julius D, Brake A, Blair L, Kunisawa R, Thorner J (1984) Isolation of the putative structural gene for the lysine–arginine-cleaving endopeptidase required for processing of yeast prepro-alpha-factor. Cell 37:1075–1089
Rehfeld JF, Goetze JP (2003) The posttranslational phase of gene expression: new possibilities in molecular diagnosis. Curr Mol Med 3:25–38
Gomord V, Faye L (2004) Posttranslational modification of therapeutic proteins in plants. Curr Opin Plant Biol 7(2):171–181
Van de Ven WJM, Roebroek A (1993) Structure and function of eukaryotic proprotein processing enzymes of the subtilisin family of serine proteases. Crit Rev Oncog 4:115–136
Routtenberg A, Rekart JL (2005) Post-translational protein modification as the substrate for long-lasting memory. Trends Neurosci 28(1):12–19
Seidah NG, Mowla SJ, Hamelin J, Mamarbachi AM, Touré BB, Benjannet S, Basak A, Munzer JS, Zhong M, Marcinkiewicz J, Barale JC, Lazure C, Murphy RA, Chrétien M, Marcinkiewicz M (1999) Mammalian subtilisin/kexin isozyme SKI-1: a widely expressed proprotein convertase with a unique cleavage specificity and cellular localization. Proc Natl Acad Sci U S A 96:1321–1326
Cheng D, Espenshade PJ, Slaughter CA, Jaen JC, Brown MS, Goldstein JL (1999) Secreted site-1 protease cleaves peptides corresponding to luminal loop of sterol regulatory element-binding proteins. J Biol Chem 274:22805–22812
Seidah NG, Benjannet S, Wickham L, Marcinkiewicz J, Jasmin SB, Stifani S, Basak A, Prat A, Chrétien M (2003) The novel secretory proprotein convertase NARC-1: potential role in liver regeneration and neuronal differentiation. Proc Natl Acad Sci U S A 100:928–933
Naureckiene S, Ma L, Sreekumar K, Purandare U, Lo CF, Huang Y, Chiang LW, Grenier JM, Ozenberger BA, Jacobsen JS, Kennedy JD, DiStefano PS, Wood A, Bingham B (2003) Functional characterization of Narc 1, a novel proteinase related to proteinase K. Arch Biochem Biophys 420:55–67
Rawlings ND, Barrett AJ (1999) MEROPS: the peptidase database. Nucleic Acids Res 27:325–331
Chrétien M, Mbikay M, Gaspar L, Seidah NG (1995) Proprotein convertases and the pathophysiology of human diseases: prospective considerations. Proc Assoc Am Physicians 107:47–66
Reeder BA, Senthilselvan A, Despres JP, Angel A, Liu L, Wang H, Rabkin SW (1997) The association of cardiovascular disease risk factors with abdominal obesity in Canada. Can Med Assoc J 157(Suppl 1):S39–S45
Rosenbaum M, Leibel RL, Hirsch J (1997) Obesity. N Engl J Med 337:396–407
Croissandeau G, Basak A, Seidah NG, Chrétien M, Mbikay M (2002) Proprotein convertases are important mediators of the adipocyte differentiation of mouse 3T3-L1 cells. J Cell Sci 115:1203–1211
Spruce MC, Potter J, Coppini DV (2003) The pathogenesis and management of painful diabetic neuropathy: a review. Diabet Med 20:88–98
Shapiro J, Sciaky N, Lee J, Bosshart H, Angeletti RH, Bonifacino JS (1997) Localization of endogenous furin in cultured cell lines. J Histochem Cytochem 45:3–12
Furuta M, Yano H, Zhou A, Rouille Y, Holst J, Carroli M, Ravazzola M, Orci L, Furuta H, Steiner DF (1997) Defective prohormone processing and altered pancreatic islet morphology in mice lacking active SPC2. Proc Natl Acad Sci U S A 94:6646–6651
Stawowy P, Meyborg H, Stibenz D, Stawowy NB, Roser M, Thanabalasingam U, Veinot JP, Chrétien M, Seidah NG, Fleck E, Graf K (2005) Furin-like proprotein convertases are central regulators of the membrane type matrix metalloproteinase–pro-matrix metalloproteinase–2 proteolytic cascade in atherosclerosis. Circulation 111:2820–2827
Khatib AM, Siegfried G, Chrétien M, Metrakos P, Seidah NG (2002) Proprotein convertases in tumor progression and malignancy: novel targets in cancer therapy. Am J Pathol 160:1921–1935
Vassar R, Bennett BD, Babu-Khan S, Kahn S, Mendiaz EA, Denis P, Teplow DB, Ross S, Amarante P, Loeloff R, Luo Y, Fisher S, Fuller J, Edenson S, Lile J, Jarosinski MA, Biere AL, Curran E, Burgess T, Louis JC, Collins F, Treanor J, Rogers G, Citron M (1999) beta-Secretase cleavage of Alzheimer’s amyloid precursor protein by the transmembrane aspartic protease BACE. Science 286:735–741
Benjannet S, Elagoz A, Wickham L, Mamarbachi M, Munzer JS, Basak A, Lazure C, Cromlish JA, Sisodia S, Checler F, Chrétien M, Seidah NG (2001) Post-translational processing of beta-secretase (beta-amyloid-converting enzyme) and its ectodomain shedding. The pro- and transmembrane/cytosolic domains affect its cellular activity and amyloid-beta production. J Biol Chem 276:10879–10887
Decroly E, Vandenbranden M, Ruysschaert JM, Jacob GS, Howard SC, Marshall G, Kompelli A, Basak A, Jean F, Lazure C, Benjannet S, Chrétien M, Day R, Seidah NG (1994) The convertases furin and PC1 can both cleave the human immunodeficiency virus (HIV)-1 envelope glycoprotein gp160 into gp120 (HIV-1 SU) and gp41 (HIV-I TM). J Biol Chem 269:12240–12247
Basak A, Zhong M, Munzer JS, Chrétien M, Seidah NG (2001) Implication of the proprotein convertases furin, PC5 and PC7 in the cleavage of surface glycoproteins of Hong Kong, Ebola and respiratory syncytial viruses: a comparative analysis with fluorogenic peptides. Biochem J 353:537–545
Molloy SS, Anderson ED, Jean F, Thomas G (1999) Bi-cycling the furin pathway: from TGN localization to pathogen activation and embryogenesis. Trends Cell Biol 9:28–35
Jean F, Stella K, Thomas L, Liu G, Xiang Y, Reason AJ, Thomas G (1998) alpha 1-Antitrypsin Portland, a bioengineered serpin highly selective for furin: application as an antipathogenic agent. Proc Natl Acad Sci U S A 95:7293–7298
Arnaoutova I, Smith AM, Coates LC, Sharpe JC, Dhanvantari S, Snell CR, Birch NP, Loh YP (2003) The prohormone processing enzyme PC3 is a lipid-raft associated transmembrane protein. Biochemistry 42:10445–10455
Bogdanovic S, Langlands B (2005) Proteases: technologies and opportunities for drug discovery. D and M D publications, Westborough, MA, USA (http://www.drugandmarket.com)
Boado RJ, Tsukamoto H, Pardridge WM (1998) Drug delivery of antisense molecules to the brain for treatment of Alzheimer’s disease and cerebral AIDS. J Pharm Sci 87:1308–1315
Hamstra DA, Rehemtulla A (1999) Toward an enzyme/prodrug strategy for cancer gene therapy: endogenous activation of carboxypeptidase A mutants by the PACE/Furin family of propeptidases. Hum Gene Ther 10:235–248
Fugere M, Day R (2002) Inhibitors of the subtilase-like pro-protein convertases (SPCs). Curr Pharm Des 8:549–562
Lazure C (2002) The peptidase zymogen proregions: nature’s way of preventing undesired activation and proteolysis. Curr Pharm Des 8:511–531
Basak A, Lazure C (2003) Synthetic peptides derived from the prosegments of proprotein convertase 1/3 and furin are potent inhibitors of both enzymes. Biochem J 373:231–239
Basak A, Jean F, Seidah NG, Lazure C (1994) Design and synthesis of novel inhibitors of prohormone convertases. Int J Pept Protein Res 44:253–261
Apletalina EV, Juliano MA, Juliano L, Lindberg I (2000) Structure–function analysis of the 7B2 CT peptide. Biochem Biophys Res Commun 267:940–942
Basak A, Lazure C (1997) Arginine and peptidyl oximes are able to inhibit both proprotein convertases PC1 and furin. Protein Pept Lett 4:187–194
Cameron A, Appel J, Houghten RA, Lindberg I (2000) Polyarginines are potent furin inhibitors. J Biol Chem 275:36741–36749
Anderson ED, Thomas L, Hayflick JS, Thomas G (1993) Inhibition of HIV-1 gp160-dependent membrane fusion by a furin-directed alpha 1-antitrypsin variant. J Biol Chem 268:24887–24891
Basak A, Schmidt C, Ismail AA, Seidah NG, Chrétien M, Lazure C (1995) Peptidyl substrates containing unnatural amino acid at the P′1 position are potent inhibitors of prohormone convertases. Int J Pept Protein Res 46:228–237
Mbikay M, Seidah NG, Chrétien M (2001) Neuroendocrine secretory protein 7B2: structure, expression and functions. Biochem J 357:329–342
Basak A, Cooper S, Roberge AG, Banik UK, Chrétien M, Seidah NG (1999) Inhibition of proprotein convertases-1, -7 and furin by diterpines of Andrographis paniculata and their succinoyl esters. Biochem J 338:107–113
Ohkubo K, Naito Y, Fujiwara T, Miyazaki JI, Ikehara Y, Ono J (2003) Inhibitory effect of the α1-antitrypsin Pittsburgh like-mutant (α1-PIM/R) on proinsulin processing in the regulated secretory pathway of the pancreatic β-cell line MIN6. Endocrinol J 50:9–20
Basak A, Dong F, Basak S (2003) In: Chorev M, Sawyer TK (eds) Peptides, peptide revolution: genomics, proteomics and therapeutics. American Peptide Society, pp 343–345
Villemure M, Fournier A, Gauthier D, Rabah N, Wilkes BC, Lazure C (2003) Barley serine proteinase inhibitor 2-derived cyclic peptides as potent and selective inhibitors of convertases PC1/3 and furin. Biochemistry 42:9659–9668
Komiyama T, Fuller RS (2000) Engineered eglin c variants inhibit yeast and human proprotein processing proteases, Kex2 and furin. Biochemistry 39:15156–15165
Boudreault A, Gauthier D, Lazure C (1998) Proprotein convertase PC1/3-related peptides are potent slow tight-binding inhibitors of murine PC1/3 and furin. J Biol Chem 273:31574–31580
Zhong M, Munzer JS, Basak A, Benjannet S, Mowla SJ, Decroly E, Chrétien M, Seidah NG (1999) The prosegments of furin and PC7 as potent inhibitors of proprotein convertases in vitro and ex vivo assessment of their efficacy and selectivity. J Biol Chem 274:33913–33920
Nour N, Basak A, Chrétien M, Seidah NG (2003) Structure–function analysis of the prosegment of the proprotein convertase PC5A. J Biol Chem 278:2886–2895
Podsiadlo P, Komiyama T, Fuller RS, Blum O (2004) Furin inhibition by compounds of copper and zinc. J Biol Chem 279:36219–36227
Brinkerhoff CJ, Podsiadlo P, Komiyama T, Fuller RS, Blum O (2002) Protease inhibitors formed in-situ from copper and tridentate chelates: a generalized approach towards metal-based pharmaceutics. ChemBioChem 3:1141–1143
Apletalina E, Appel J, Lamango NS, Houghten RA, Lindberg I (1998) Identification of inhibitors of prohormone convertases 1 and 2 using a peptide combinatorial library. J Biol Chem 273:26589–26595
Basak A, Ernst B, Brewer D, Seidah NG, Munzer JS, Lazure C, Lajoie GA (1997) Histidine-rich human salivary peptides are inhibitors of proprotein convertases furin and PC7 but act as substrate for PC1. J Pept Res 49:596–603
Jean F, Basak A, Dugas H, Seidah NG, Chrétien M, Lazure C (1994) Peptides: chemistry structure and biology. In: Hodges RS, Smith JA (eds) Proceedings of 13th American Peptide Symposium. ESCOM, Leiden, The Netherlands, pp 613–615
Jean F, Basak A, Rondeau N, Benjannet S, Hendy GN, Seidah NG, Chrétien M, Lazure C (1993) Enzymic characterization of murine and human prohormone convertase-1 (mPC1 and hPC1) expressed in mammalian GH4C1 cells. Biochem J 292:891–899
Angliker H (1995) Synthesis of tight binding inhibitors and their action on the proprotein-processing enzyme furin. J Med Chem 38:4014–4018
Hallenberger S, Bosch V, Angliker H, Shaw E, Klenk HD, Garten W (1992) Inhibition of furin-mediated cleavage activation of HIV-1 glycoprotein gpl60. Nature 360:358–361
Fricker LD, McKinzie AA, Sun J, Curran E, Qian Y, Yan L, Patterson SD, Courchesne PL, Richards B, Levin N, Mzhavia N, Devi LA, Douglass J (2000) Identification and characterization of proSAAS, a granin-like neuroendocrine peptide precursor that inhibits prohormone processing. J Neurosci 20:639–648
Basak A, Koch P, Dupelle M, Fricker LD, Devi LA, Chrétien M, Seidah NG (2001) Inhibitory specificity and potency of proSAAS-derived peptides toward proprotein convertase 1. J Biol Chem 276:32720–32728
Dahlen JR, Jean F, Thomas G, Foster DC, Kisiel W (1998) Inhibition of soluble recombinant furin by human proteinase inhibitor 8. J Biol Chem 273:1851–1854
Cornwall GA, Cameron A, Lindberg I, Hardy DM, Cormier N, Hsia N (2003) CRES inhibits the serine protease prohormone convertase 2. Endocrinology 144:901–908
Lu W, Zhang W, Molloy SS, Thomas G, Ryan K, Chiang Y, Anderson S, Laskowski M Jr (1993) Arg15–Lys17–Arg18 turkey ovomucoid third domain inhibits human furin. J Biol Chem 268:14583–14585
Kan CC, Solomon E, Belt KT, Chain AC, Hiorns LR, Fey G (1985) Nucleotide sequence of cDNA encoding human alpha 2-macroglobulin and assignment of the chromosomal locus. Proc Natl Acad Sci U S A 82:2282–2286
Van Rompaey L, Ayoubi T, Van De Ven W, Marynen P (1997) Inhibition of intracellular proteolytic processing of soluble proproteins by an engineered α2-macroglobulin containing a furin recognition sequence in the bait region. Biochem J 326:507–514
Hipler K, Priestle JP, Rahuel J, Grutter MG (1996) Active site binding loop stabilization in the subtilisin inhibitor eglin c: structural and functional studies on specifically designed mutants in complex with subtilisin and the uncomplexed inhibitor. Adv Exp Med Biol 379:43–47
Fu X, Inouye M, Shinde U (2000) Folding pathway mediated by an intramolecular chaperone. The inhibitory and chaperone functions of the subtilisin propeptide are not obligatorily linked. J Biol Chem 275:16871–16878
Basak A, Koch P, Dupelle M, Sirois F, Chrétien M, Seidah NG, Mbikay M (2001) In: Lebl M, Houghten RA (eds) Peptides, the wave of the future. Kluwer Academic Publishers, Dordrecht, pp 558–560
Bahbouhi B, Chazal N, Seidah NG, Chiva C, Kogan M, Albericio F, Giralt E, Bahraoui E (2002) Effects of l- and d-REKR amino acid-containing peptides on HIV and SIV envelope glycoprotein precursor maturation and HIV and SIV replication. Biochem J 366:863–872
Basak A, Touré BB, Lazure C, Mbikay M, Chrétien M, Seidah NG (1999) Enzymic characterization in vitro of recombinant proprotein convertase PC4. Biochem J 343:29–37
Jean F, Boudreault A, Basak A, Seidah NG, Lazure C (1995) Fluorescent peptidyl substrates as an aid in studying the substrate specificity of human prohormone convertase PC1 and human furin and designing a potent irreversible inhibitor. J Biol Chem 270:19225–19231
Maquoi E, Noel A, Frankenne F, Angliker H, Murphy G, Foidart JM (1998) Inhibition of matrix metalloproteinase 2 maturation and HT1080 invasiveness by a synthetic furin inhibitor. FEBS Lett 424:262–266
Milner JM, Rowan AD, Elliott SF, Cawston TE (2003) Inhibition of furin-like enzymes blocks interleukin-1alpha/oncostatin M-stimulated cartilage degradation. Arthritis Rheum 48:1057–1066
Denault JB, D’Orleans-Juste P, Masaki T, Leduc R (1995) Inhibition of convertase-related processing of proendothelin-1. J Cardiovasc Pharmacol 26(Suppl 3):S47–S50
Pearton DJ, Nirunsuksiri W, Rehemtulla A, Lewis SP, Presland RB, Dale BA (2001) Convertases expression and localization in epidermis: evidence for multiple roles and substrates. Exp Dermatol 10:193–203
Qiu Q, Basak A, Mbikay M, Tsang BK, Gruolin A (2005) Role of pro-IGF-II processing by proptrotein convertrase 4 in human placentral development. Proc Natl Acad Sci U S A 102:11047–11052
Basak A, Lotfipour F (2005) Modulating furin activity with designed mini-PDX peptides: Synthesis and in vitro evaluation. FEBS Lett 579:4813–4821
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The work was supported by grants from CFI, CIHR, NSERC and the University of Ottawa.
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Basak, A. Inhibitors of proprotein convertases. J Mol Med 83, 844–855 (2005). https://doi.org/10.1007/s00109-005-0710-0
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DOI: https://doi.org/10.1007/s00109-005-0710-0