Expression of a human proprotein processing enzyme: correct cleavage of the von Willebrand factor precursor at a paired basic amino acid site.

Wise, Robert J.; Barr, Philip J.; Wong, P A; Kiefer, Michael; Brake, Anthony J.; Kaufman, Randal J.

doi:10.1073/pnas.87.23.9378

Cited by 285 publications

(184 citation statements)

References 47 publications

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“…The mammalian members of the prohormone convertase family include furin, also known as PACE [2][3][4][5][6] ; PC2 [7,8] ; PC1, also identified as PC3 [8][9][10][11] ; PC4 [12,13] ; PC5, also known as PC6 [14][15][16] ; PACE4 [17], also identified as PC7 [18] ; and most recently PC8, also referred to as LPC, rPC7 and SPC7 [19][20][21][22]. Furin, PACE4, PC5\6 and PC8 are expressed in a wide variety of cell lines and tissues, although variation in the relative abundance of these enzymes is observed [6,14,15,17,19,21,23].…”

Section: Introductionmentioning

confidence: 99%

Gene organization and alternative splicing of human prohormone convertase PC8

Goodge

Thomas

Martın

et al. 1998

Biochemical Journal

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The mammalian Ca2+-dependent serine protease prohormone convertase PC8 is expressed ubiquitously, being transcribed as 3.5, 4.3 and 6.0 kb mRNA isoforms in various tissues. To determine the origin of these various mRNA isoforms we report the characterization of the human PC8 gene, which has been previously localized to chromosome 11q23–24. Consisting of 16 exons, the human PC8 gene spans approx. 27 kb. A comparison of the position of intron–exon junctions of the human PC8 gene with the gene structures of previously reported prohormone convertase genes demonstrated a divergence of the human PC8 from the highly conserved nature of the gene organization of this enzyme family. The nucleotide sequence of the 5´-flanking region of the human PC8 is reported and possesses putative promoter elements characteristic of a GC-rich promoter. Further supporting the potential role of a GC-rich promoter element, multiple transcriptional initiation sites within a 200 bp region were demonstrated. We propose that the various mRNA isoforms of PC8 result from the inclusion of intronic sequences within transcripts.

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Section: Introductionmentioning

confidence: 99%

Gene organization and alternative splicing of human prohormone convertase PC8

Goodge

Thomas

Martın

et al. 1998

Biochemical Journal

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“…Here, r-vWF-producing CHO cells were additionally manipulated to co-express the recombinant subtilisinlike proprotein processing enzyme Furin (r-Furin) [9,10] in order to ensure complete processing of pro-vWF. Recombinant CHO cells (cell clone # 904) were cultivated for the first time on an industrial scale in high cell density perfusion bio-reactors.…”

Section: : Introductionmentioning

confidence: 99%

Structural analysis of recombinant von Willebrand factor produced at industrial scale fermentation of transformed CHO cells co‐expressing recombinant furin

Fischer¹,

Schlokat²,

Mitterer³

et al. 1995

FEBS Letters

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“…To assess the enzymic activities of the mutants, each was cotransfected into furin-deficient CHO RPE.40 cells along with a cDNA encoding vWf ( Figure 4A), a well-documented furin substrate [27,28]. Basal processing of the vWf precursor was observed following pulse labelling of cells transfected with provWf alone.…”

Section: Processing Activity Of Pro-region Mutantsmentioning

confidence: 99%

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

Bissonnette

Charest

Longpré

et al. 2004

Biochemical Journal

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The pro-region of the subtilisin-like convertase furin acts early in the biosynthetic pathway as an intramolecular chaperone to enable proper folding of the zymogen, and later on as an inhibitor to constrain the activity of the enzyme until it reaches the transGolgi network. To identify residues that are important for proregion function, we initially identified amino acids that are conserved among the pro-regions of various mammalian convertases. Site-directed mutagenesis of 17 selected amino acids within the 89-residue pro-region and biosynthetic labelling revealed that I60A-furin and H66A-furin were rapidly degraded in a proteasome-dependent manner, while W34A-furin and F67A-furin did not show any autocatalytic activation. Intriguingly, the latter mutants proteolytically cleaved pro-von Willebrand factor precursor to the mature polypeptide, suggesting that the mutations permitted proper folding, but did not allow the pro-region to exercise its role in inhibiting the enzyme. Homology modelling of furin's pro-region revealed that residues Ile-60 and His-66 might be crucial in forming the binding interface with the catalytic domain, while residues Trp-34 and Phe-67 might be involved in maintaining a hydrophobic core within the pro-region itself. These results provide structural insights into the dual role of furin's proregion.

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Expression of a human proprotein processing enzyme: correct cleavage of the von Willebrand factor precursor at a paired basic amino acid site.

Cited by 285 publications

References 47 publications

Gene organization and alternative splicing of human prohormone convertase PC8

Gene organization and alternative splicing of human prohormone convertase PC8

Structural analysis of recombinant von Willebrand factor produced at industrial scale fermentation of transformed CHO cells co‐expressing recombinant furin

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

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