Identification of furin pro-region determinants involved in folding and activation

Bissonnette, Lyne; Charest, Gabriel; Longpré, Jean‐Michel; Lavigne, Pierre; Leduc, Richard

doi:10.1042/bj20031902

Cited by 18 publications

(12 citation statements)

References 40 publications

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“…Recombinant enzymes lacking different sequences between the pro-and the metalloproteinase domains were efficiently synthesized and secreted but exhibited no activity. This result suggests that the pro-domain is not only responsible for the repression of enzyme activity but is also essential for the correct folding of the protein as observed for other enzymes (28).…”

Section: Discussionmentioning

confidence: 55%

Domains and Maturation Processes That Regulate the Activity of ADAMTS-2, a Metalloproteinase Cleaving the Aminopropeptide of Fibrillar Procollagens Types I–III and V

Colige

Ruggiero

Vandenberghe

et al. 2005

Journal of Biological Chemistry

108

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Processing of fibrillar collagens is required to generate collagen monomers able to self-assemble into elongated and cylindrical collagen fibrils. ADAMTS-2 belongs to the "A disintegrin and metalloproteinase with thrombospondin type 1 motifs" (ADAMTS) family. It is responsible for most of the processing of the aminopropeptide of type I procollagen in the skin, and it also cleaves type II and type III procollagens. ADAMTS are complex secreted enzymes that are implicated in various physiological and pathological processes. Despite accumulating evidence indicating that their activity is regulated by ancillary domains, additional information is required for a better understanding of the specific function of each domain. We have generated 17 different recombinant forms of bovine ADAMTS-2 and characterized their processing, activity, and cleavage specificity. The results indicated the following: (i) activation of the ADAMTS-2 zymogen involves several cleavages, by proprotein convertases and C-terminal processing, and generates at least seven distinct processed forms; (ii) the C-terminal domain negatively regulates enzyme activity, whereas two thrombospondin type 1 repeats are enhancer regulators; (iii) the 104-kDa form displays the highest aminoprocollagen peptidase activity on procollagen type I; (iv) ADAMTS-2 processes the aminopropeptide of ␣1 type V procollagen homotrimer at the end of the variable domain; and (v) the cleaved sequence (PA) is different from the previously described sites ((P/A)Q) for ADAMTS-2, redefining its cleavage specificity. This finding and the existence of multiple processed forms of ADAMTS-2 strongly suggest that ADAMTS-2 may be involved in function(s) other than processing of fibrillar procollagen types I-III.Collagens types I-III are synthesized as precursors (procollagens) formed by a central triple helical collagen domain extended by propeptides at both extremities. Removal of the C-propeptide, 3 by BMP-1 and related enzymes, and the N-propeptide, by aminoprocollagen peptidases (ADAMTS-2, -3, or -14), is required to generate collagen monomers able to assemble spontaneously into elongated and cylindrical collagen fibrils. It has been shown in vitro and in vivo that ADAMTS-2 is the enzyme responsible for most of the aminoprocollagen type I processing in the skin (1-3), although it was suggested that an essential function of ADAMTS-3 is the processing of aminoprocollagen type II in cartilage (4). A closely related enzyme, ADAMTS-14, also displays aminoprocollagen peptidase activity (5). More recently, it has been shown that ADAMTS-2 is also able to process aminoprocollagen type III in vitro (6). Type V collagen is a minor fibrillar collagen. Among the four different individual collagen type V chains, ␣1 is the most abundant and ubiquitous and can be found as a homotrimer or as a heterotrimer in association with type V or type XI ␣ chains (7,8). Its maturation from procollagen to collagen is reported to be different from what is observed for procollagens I-III, because the C-propeptide can...

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Section: Discussionmentioning

confidence: 55%

Domains and Maturation Processes That Regulate the Activity of ADAMTS-2, a Metalloproteinase Cleaving the Aminopropeptide of Fibrillar Procollagens Types I–III and V

Colige

Ruggiero

Vandenberghe

et al. 2005

Journal of Biological Chemistry

108

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“…68,69 According to sequence identity, the pro-domains of the mammalian PCs might not resemble each other as much as do their catalytic and P domains. Recent structure-based PC alignments 65 as well as homology modelling and mutation experiments on furin 70 show, however, that these pro-domains seem to exhibit a similar overall topology. In the initial pro-PCs, the pro-domains might cap the N termini of helices Ca3 and Ca4 (see Figures 1(a) and 3), presumably similar to the interaction of the subtilisin pro-peptides and the Figure 3.…”

Section: Overall Structure Of the P Domainsmentioning

confidence: 83%

Proprotein Convertase Models based on the Crystal Structures of Furin and Kexin: Explanation of their Specificity

Henrich

Lindberg

Bode

et al. 2005

Journal of Molecular Biology

142

161

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“…In addition, in vitro activation of PC1, which contains the evolutionarily conserved His residue corresponding to His 69 of furin, also requires an acidic pH for activation. 3 However, a recent study demonstrates that an H69A mutation had no effect on the ability of overexpressed furin to cleave pro-von Willebrand factor in 293 cells (39). This discrepancy between the H69L and H69A substitutions may be attributed to a larger van der Waals volume of Leu, which may result in a more prominent effect on cleavage and activation because of stronger hydrophobic interactions, differences in the assays used to detect furin activity, or perhaps differences in the cell lines used (10,11).…”

Section: Discussionmentioning

confidence: 99%

Identification of a pH Sensor in the Furin Propeptide That Regulates Enzyme Activation

Féliciangéli

Thomas

Scott

et al. 2006

Journal of Biological Chemistry

102

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The folding and activation of furin occur through two pH-and compartment-specific autoproteolytic steps. In the endoplasmic reticulum (ER), profurin folds under the guidance of its prodomain and undergoes an autoproteolytic excision at the consensus furin site Arg-Thr-Lys-Arg 107 2 generating an enzymatically masked furin-propeptide complex competent for transport to late secretory compartments. In the mildly acidic environment of the trans-Golgi network/endosomal system, the bound propeptide is cleaved at the internal site 69 HRGVTKR 75 2, unmasking active furin capable of cleaving substrates in trans. Here, by using cellular, biochemical, and modeling studies, we demonstrate that the conserved His 69 is a pH sensor that regulates the compartment-specific cleavages of the propeptide. In the ER, unprotonated His 69 stabilizes a solvent-accessible hydrophobic pocket necessary for autoproteolytic excision at Arg 107 . Profurin molecules unable to form the hydrophobic pocket, and hence, the furin-propeptide complex, are restricted to the ER by a PACS-2-and COPI-dependent mechanism. Once exposed to the acidic pH of the late secretory pathway, protonated His 69 disrupts the hydrophobic pocket, resulting in exposure and cleavage of the internal cleavage site at Arg 75 to unmask the enzyme. Together, our data explain the pH-regulated activation of furin and how this His-dependent regulatory mechanism is a model for other proteins.The pH gradient formed by the various membranous compartments comprising the secretory and endocytic pathways has essential and manifold roles ranging from the regulation of protein traffic to the control of protein conformation and enzyme activities, including the processing of prohormones and proproteins by proprotein convertases (PCs). 2 The PCs, a family of calcium-dependent serine endoproteases, cleave proproteins and prohormones at doublets or clusters of basic amino acids thus generating mature bioactive proteins as well as hormonal processing intermediates that require additional post-translational modifications to gain full bioactivity (1, 2). The proteolytic maturation of prohormones by the neuroendocrine-specific PC1/3 and PC2 requires the acidic pH of maturing secretory granules (3-5), whereas the proteolytic activation of proprotein substrates by the ubiquitously expressed PC furin in the trans-Golgi network (TGN)/endosomal system is compartment-and hence pH-specific (2, 6). This compartmentspecific processing by furin is mediated in part by the pH-dependent changes in the conformation of furin substrates as well as the utilization of pH-sensitive furin cleavage sites. Furin efficiently cleaves a number of proprotein substrates at the consensus site -Arg-X-Lys/Arg-Arg-2 (2). Kinetic studies show that the P1 and P4 Arg residues are required for the efficient processing of furin substrates, whereas the P2 Arg has a modulatory role (7,8). However, at acidic pH the absence of a P2 or P4 Arg can be compensated for by the presence of positively charged residues at P6 or possibly adjacen...

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Identification of furin pro-region determinants involved in folding and activation

Cited by 18 publications

References 40 publications

Domains and Maturation Processes That Regulate the Activity of ADAMTS-2, a Metalloproteinase Cleaving the Aminopropeptide of Fibrillar Procollagens Types I–III and V

Domains and Maturation Processes That Regulate the Activity of ADAMTS-2, a Metalloproteinase Cleaving the Aminopropeptide of Fibrillar Procollagens Types I–III and V

Proprotein Convertase Models based on the Crystal Structures of Furin and Kexin: Explanation of their Specificity

Identification of a pH Sensor in the Furin Propeptide That Regulates Enzyme Activation

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