Evidence for furin-type activity-mediated C-terminal processing of profibrillin-1 and interference in the processing by certain mutations.

Fibrillin-1 is a major component of the 10 nm microfibrils of the extracellular matrix (ECM). It is synthesized as an approximately 350 kDa precursor molecule, profibrillin-1, which is proteolytically processed into its biologically active approximately 320 kDa form. Furin, a calcium-dependent endoprotease of the subtilisin family, which is known to be the processing enzyme for a variety of proproteins, is believed to be responsible for the N-terminal proteolytic cleavage of profibrillin-1. In this article we provide several lines of evidence that the C-terminal trimming of profibrillin-1 also occurs via a furin-type activity. Edman degradation of a small recombinant C-terminal subdomain of fibrillin-1 revealed complete processing of the peptide immediately after the tribasic recognition sequence (R-X-K/R-R) for furin. In vitro expression experiments using another recombinant construct consisting of the C-terminal half of fibrillin-1 indicated that disruption of the putative recognition sequence for furin by site-directed mutagenesis drastically impairs proteolytic processing of the propeptide. In addition, our results suggest that the N-terminal half of fibrillin-1 is necessary for its incorporation into the ECM.

[1]  D. Keene,et al.  Fibrillin-1: organization in microfibrils and structural properties. , 1996, Journal of molecular biology.

[2]  R. Leduc,et al.  Processing of Transforming Growth Factor 1 Precursor by Human Furin Convertase (*) , 1995, The Journal of Biological Chemistry.

[3]  D. Milewicz,et al.  A mutation in FBN1 disrupts profibrillin processing and results in isolated skeletal features of the Marfan syndrome. , 1995, The Journal of clinical investigation.

[4]  E. Smiley,et al.  Primary Structure and Developmental Expression of Fbn-1, the Mouse Fibrillin Gene (*) , 1995, The Journal of Biological Chemistry.

[5]  R. Roth,et al.  Accurate and efficient cleavage of the human insulin proreceptor by the human proprotein-processing protease furin. Characterization and kinetic parameters using the purified, secreted soluble protease expressed by a recombinant baculovirus. , 1994, The Journal of biological chemistry.

[6]  J. Womack,et al.  Sequence of the coding region of the bovine fibrillin cDNA and localization to bovine chromosome 10. , 1994, Genomics.

[7]  L. Peltonen,et al.  Analyses of truncated fibrillin caused by a 366 bp deletion in the FBN1 gene resulting in Marfan syndrome. , 1994, The Biochemical journal.

[8]  R. Mecham,et al.  Structure and expression of fibrillin-2, a novel microfibrillar component preferentially located in elastic matrices , 1994, The Journal of cell biology.

[9]  G. Thomas,et al.  Intracellular trafficking and activation of the furin proprotein convertase: localization to the TGN and recycling from the cell surface. , 1994, The EMBO journal.

[10]  C. Hayward,et al.  Identification of a novel nonsense mutation in the fibrillin gene (FBN1) using nonisotopic techniques , 1994, Human mutation.

[11]  M. Komada,et al.  Proteolytic processing of the hepatocyte growth factor/scatter factor receptor by furin , 1993, FEBS letters.

[12]  H. Dietz,et al.  Fibrillin binds calcium and is coded by cDNAs that reveal a multidomain structure and alternatively spliced exons at the 5' end. , 1993, Genomics.

[13]  R. Timpl,et al.  A single EGF‐like motif of laminin is responsible for high affinity nidogen binding. , 1993, The EMBO journal.

[14]  D. Steiner,et al.  The new enzymology of precursor processing endoproteases. , 1992, The Journal of biological chemistry.

[15]  A. Rehemtulla,et al.  Regulation of PACE propeptide-processing activity: requirement for a post-endoplasmic reticulum compartment and autoproteolytic activation. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[16]  K. Klimpel,et al.  Human furin is a calcium-dependent serine endoprotease that recognizes the sequence Arg-X-X-Arg and efficiently cleaves anthrax toxin protective antigen. , 1992, The Journal of biological chemistry.

[17]  S. Takahashi,et al.  Purification and characterization of furin, a Kex2-like processing endoprotease, produced in Chinese hamster ovary cells. , 1992, The Journal of biological chemistry.

[18]  A. Rehemtulla,et al.  Preferred sequence requirements for cleavage of pro-von Willebrand factor by propeptide-processing enzymes. , 1992, Blood.

[19]  H. Wiedemann,et al.  Recombinant nidogen consists of three globular domains and mediates binding of laminin to collagen type IV. , 1991, The EMBO journal.

[20]  K. Nakayama,et al.  Structure and expression of mouse furin, a yeast Kex2-related protease. Lack of processing of coexpressed prorenin in GH4C1 cells. , 1990, The Journal of biological chemistry.

[21]  A. Brake,et al.  Expression of a human proprotein processing enzyme: correct cleavage of the von Willebrand factor precursor at a paired basic amino acid site. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[22]  R. Pyeritz,et al.  Immunohistologic abnormalities of the microfibrillar-fiber system in the Marfan syndrome. , 1990, The New England journal of medicine.

[23]  C. Gorman,et al.  Transient production of proteins using an adenovirus transformed cell line , 1990 .

[24]  S. Kawabata,et al.  Factor IX Kawachinagano: impaired function of the Gla‐domain caused by attached propeptide region due to substitution of arginine by glutamine at position −4 , 1989, British journal of haematology.

[25]  A. Kosaki,et al.  Insulin-resistant diabetes due to a point mutation that prevents insulin proreceptor processing. , 1988, Science.

[26]  R. Webster,et al.  Sequence requirements for cleavage activation of influenza virus hemagglutinin expressed in mammalian cells. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[27]  E. Engvall,et al.  Fibrillin, a new 350-kD glycoprotein, is a component of extracellular microfibrils , 1986, The Journal of cell biology.

[28]  D. Rees,et al.  Defective propeptide processing of blood clotting factor IX caused by mutation of arginine to glutamine at position −4 , 1986, Cell.

[29]  H. Luthman,et al.  High efficiency polyoma DNA transfection of chloroquine treated cells. , 1983, Nucleic acids research.

[30]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.