Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
Differences in the autocatalytic cleavage of pro-PC2 and pro-PC3 can be attributed to sequences within the propeptide and Asp310 of pro-PC2.
Scougall K
,
Taylor NA
,
Jermany JL
,
Docherty K
,
Shennan KI
.
???displayArticle.abstract??? PC2 and PC3 are subtilisin-like proteases involved in the maturation of prohormones and proneuropeptides within neuroendocrine cells. They are synthesized as zymogens that undergo autocatalytic maturation within the secretory pathway. Maturation of pro-PC2 is slow (t12 >8 h), exhibits a pH optimum of 5.5 and is dependent on calcium (K0.5 2 mM), while pro-PC3 maturation is relatively rapid (t12 15 min), exhibits a neutral pH optimum and is not calcium dependent. These differences in the rates and optimal conditions for activation of the proteases may contribute to the diversity of products generated by these proteases in different cell types. Although highly similar, there are two major differences between pro-PC2 and pro-PC3: the presence of an aspartate at position 310 in pro-PC2 compared with asparagine at the equivalent position in pro-PC3 (and all other members of the subtilisin family), and the N-terminal propeptides, which exhibit low sequence identity (30%). With a view to establishing the structural features that might be responsible for these differences in the maturation of pro-PC2 and pro-PC3, Asp310 in pro-PC2 was mutated to Asn, and Asn309 in pro-PC3 was mutated to Asp. Chimaeric proteins were also made consisting of the pro-region of PC2 fused to the mature portion of PC3 and the pro-region of PC3 fused to the mature region of PC2. The wild-type and mutant DNA constructs were then transcribed and translated in an in vitro system capable of supporting maturation of pro-PC2 and pro-PC3. The results demonstrated that Asp310 of pro-PC2 is responsible for the acidic pH optimum for maturation. Thus changing Asp310 to Asn shifted the pH optimum for maturation to pH 7.0. However, changing Asn309 of pro-PC3 to Asp had no effect on the optimum pH for maturation of pro-PC3. A chimaeric construct containing the propeptide of pro-PC2 attached to PC3 shifted the pH optimum for maturation from pH 7.0 to 6.0 and slowed down the rate of maturation (t12 >8 h). When attached to PC2, the pro-region of pro-PC3 had no effect on the optimum pH for maturation (pH 5.5-6.0), but it did accelerate the rate of maturation (t12 2 h). These results demonstrate that Asp310 and the pro-region of pro-PC2 contribute to the acidic pH optimum and low rate of maturation of this zymogen relative to its closely related homologue PC3.
Anderson,
Activation of the furin endoprotease is a multiple-step process: requirements for acidification and internal propeptide cleavage.
1997, Pubmed
Anderson,
Activation of the furin endoprotease is a multiple-step process: requirements for acidification and internal propeptide cleavage.
1997,
Pubmed
Anderson,
A view of acidic intracellular compartments.
1988,
Pubmed
Barr,
cDNA and gene structure for a human subtilisin-like protease with cleavage specificity for paired basic amino acid residues.
1991,
Pubmed
Benjannet,
7B2 is a specific intracellular binding protein of the prohormone convertase PC2.
1995,
Pubmed
Benjannet,
Proprotein conversion is determined by a multiplicity of factors including convertase processing, substrate specificity, and intracellular environment. Cell type-specific processing of human prorenin by the convertase PC1.
1992,
Pubmed
Benjannet,
Comparative biosynthesis, covalent post-translational modifications and efficiency of prosegment cleavage of the prohormone convertases PC1 and PC2: glycosylation, sulphation and identification of the intracellular site of prosegment cleavage of PC1 and PC2.
1993,
Pubmed
Benjannet,
Structure-function studies on the biosynthesis and bioactivity of the precursor convertase PC2 and the formation of the PC2/7B2 complex.
1995,
Pubmed
Braks,
7B2 is a neuroendocrine chaperone that transiently interacts with prohormone convertase PC2 in the secretory pathway.
1994,
Pubmed
,
Xenbase
Bruzzaniti,
PC8 [corrected], a new member of the convertase family.
1996,
Pubmed
Chaudhuri,
The neuroendocrine protein 7B2 acts as a molecular chaperone in the in vitro folding of human insulin-like growth factor-1 secreted from yeast.
1995,
Pubmed
Coates,
Posttranslational maturation of the prohormone convertase SPC3 in vitro.
1997,
Pubmed
Creemers,
Identification of a transferable sorting domain for the regulated pathway in the prohormone convertase PC2.
1996,
Pubmed
Dupuy,
Processing of prodynorphin by the prohormone convertase PC1 results in high molecular weight intermediate forms. Cleavage at a single arginine residue.
1994,
Pubmed
Eder,
Pro-sequence-assisted protein folding.
1995,
Pubmed
Fuller,
Yeast prohormone processing enzyme (KEX2 gene product) is a Ca2+-dependent serine protease.
1989,
Pubmed
Goodman,
Autoproteolytic activation of the mouse prohormone convertase mPC1.
1994,
Pubmed
Halban,
Sorting and processing of secretory proteins.
1994,
Pubmed
Kiefer,
Identification of a second human subtilisin-like protease gene in the fes/fps region of chromosome 15.
1991,
Pubmed
Krieg,
Functional messenger RNAs are produced by SP6 in vitro transcription of cloned cDNAs.
1984,
Pubmed
,
Xenbase
Leduc,
Activation of human furin precursor processing endoprotease occurs by an intramolecular autoproteolytic cleavage.
1992,
Pubmed
Lindberg,
Evidence for cleavage of the PC1/PC3 pro-segment in the endoplasmic reticulum.
1994,
Pubmed
Lusson,
cDNA structure of the mouse and rat subtilisin/kexin-like PC5: a candidate proprotein convertase expressed in endocrine and nonendocrine cells.
1993,
Pubmed
Matthews,
A highly efficient, cell-free translation/translocation system prepared from Xenopus eggs.
1991,
Pubmed
,
Xenbase
Matthews,
Autocatalytic maturation of the prohormone convertase PC2.
1994,
Pubmed
,
Xenbase
Meerabux,
A new member of the proprotein convertase gene family (LPC) is located at a chromosome translocation breakpoint in lymphomas.
1996,
Pubmed
Mizuno,
Yeast KEX2 genes encodes an endopeptidase homologous to subtilisin-like serine proteases.
1988,
Pubmed
Molloy,
Intracellular trafficking and activation of the furin proprotein convertase: localization to the TGN and recycling from the cell surface.
1994,
Pubmed
Muller,
Mechanism of the facilitation of PC2 maturation by 7B2: involvement in ProPC2 transport and activation but not folding.
1997,
Pubmed
Nakagawa,
Identification and functional expression of a new member of the mammalian Kex2-like processing endoprotease family: its striking structural similarity to PACE4.
1993,
Pubmed
Nakayama,
Identification of the fourth member of the mammalian endoprotease family homologous to the yeast Kex2 protease. Its testis-specific expression.
1992,
Pubmed
Rhodes,
What beta-cell defect could lead to hyperproinsulinemia in NIDDM? Some clues from recent advances made in understanding the proinsulin-processing mechanism.
1994,
Pubmed
Roebroek,
Evolutionary conserved close linkage of the c-fes/fps proto-oncogene and genetic sequences encoding a receptor-like protein.
1986,
Pubmed
Rufaut,
Purification and characterization of the candidate prohormone-processing enzyme SPC3 produced in a mouse L cell line.
1993,
Pubmed
Seidah,
cDNA structure, tissue distribution, and chromosomal localization of rat PC7, a novel mammalian proprotein convertase closest to yeast kexin-like proteinases.
1996,
Pubmed
Seidah,
cDNA sequence of two distinct pituitary proteins homologous to Kex2 and furin gene products: tissue-specific mRNAs encoding candidates for pro-hormone processing proteinases.
1990,
Pubmed
Seidah,
Cloning and primary sequence of a mouse candidate prohormone convertase PC1 homologous to PC2, Furin, and Kex2: distinct chromosomal localization and messenger RNA distribution in brain and pituitary compared to PC2.
1991,
Pubmed
Shen,
Biosynthesis of the prohormone convertase PC2 in Chinese hamster ovary cells and in rat insulinoma cells.
1993,
Pubmed
Shennan,
Site-directed mutagenesis and expression of PC2 in microinjected Xenopus oocytes.
1991,
Pubmed
,
Xenbase
Shennan,
Calcium- and pH-dependent aggregation and membrane association of the precursor of the prohormone convertase PC2.
1994,
Pubmed
,
Xenbase
Shennan,
Differences in pH optima and calcium requirements for maturation of the prohormone convertases PC2 and PC3 indicates different intracellular locations for these events.
1995,
Pubmed
,
Xenbase
Shinde,
Folding pathway mediated by an intramolecular chaperone.
1993,
Pubmed
Shinde,
The structural and functional organization of intramolecular chaperones: the N-terminal propeptides which mediate protein folding.
1994,
Pubmed
Siezen,
Homology modelling and protein engineering strategy of subtilases, the family of subtilisin-like serine proteinases.
1991,
Pubmed
Silen,
The alpha-lytic protease pro-region does not require a physical linkage to activate the protease domain in vivo.
1989,
Pubmed
Smeekens,
Identification of a human insulinoma cDNA encoding a novel mammalian protein structurally related to the yeast dibasic processing protease Kex2.
1990,
Pubmed
Smeekens,
Identification of a cDNA encoding a second putative prohormone convertase related to PC2 in AtT20 cells and islets of Langerhans.
1991,
Pubmed
Taylor,
Mutations within the propeptide, the primary cleavage site or the catalytic site, or deletion of C-terminal sequences, prevents secretion of proPC2 from transfected COS-7 cells.
1997,
Pubmed
Urbé,
pH-dependent processing of secretogranin II by the endopeptidase PC2 in isolated immature secretory granules.
1997,
Pubmed
Vindrola,
Biosynthesis of the prohormone convertase mPC1 in AtT-20 cells.
1992,
Pubmed
Zhou,
Structural elements that direct specific processing of different mammalian subtilisin-like prohormone convertases.
1995,
Pubmed
Zhou,
Purification and characterization of the prohormone convertase PC1(PC3).
1993,
Pubmed
Zhu,
7B2 facilitates the maturation of proPC2 in neuroendocrine cells and is required for the expression of enzymatic activity.
1995,
Pubmed
,
Xenbase
van den Ouweland,
Structural homology between the human fur gene product and the subtilisin-like protease encoded by yeast KEX2.
1990,
Pubmed
