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Biochem J. Mar 15, 1996; 314(Pt 3): 951–960.
PMCID: PMC1217149

Cellular processing of the nerve growth factor precursor by the mammalian pro-protein convertases.


In order to define the enzymes responsible for the maturation of the precursor of nerve growth factor (proNGF), its biosynthesis and intracellular processing by the pro-protein convertases furin, PC1, PC2, PACE4, PC5 and the PC5 isoform PC5/6-B were analysed using the vaccinia virus expression system in cells containing a regulated and/or a constitutive secretory pathway. Results demonstrate that in both cell types furin, and to a lesser extent PACE4 and PC5/6-B, are the best candidate proNGF convertases. Furthermore, two processed NGF forms of 16.5 and 13.5 kDa were evident in constitutively secreting cell lines such as LoVo and BSC40 cells, whereas only the 13.5 kDa form was observed in AtT20 cells, which contain secretory granules. Both forms display the same N-terminal sequence as mature NGF, and were also produced following site-directed mutagenesis of the C-terminal Arg-Arg sequence of NGF into Ala-Ala, suggesting that the difference between them is not at the C-terminus. Co-expression of proNGF with furin and either chromogranin B or secretogranin II (but not chromogranin A) in BSC40 cells eliminated the 16.5 kDa form. Data also show that N-glycosylation of the pro-segment of proNGF and trimming of the oligosaccharide chains are necessary for the exit of this precursor from the endoplasmic reticulum and its eventual processing and secretion. Sulphate labelling experiments demonstrated that proNGF is processed into mature NGF following the arrival of the precursor in the trans-Golgi network. This comparative study shows that the three candidate mammalian subtilisin/kexin-like convertases identified process proNGF into NGF and that the nature of the final processed products is dependent on the intracellular environment.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Levi-Montalcini R. The nerve growth factor 35 years later. Science. 1987 Sep 4;237(4819):1154–1162. [PubMed]
  • Angeletti RH, Hermodson MA, Bradshaw RA. Amino acid sequences of mouse 2.5S nerve growth factor. II. Isolation and characterization of the thermolytic and peptic peptides and the complete covalent structure. Biochemistry. 1973 Jan 2;12(1):100–115. [PubMed]
  • Scott J, Selby M, Urdea M, Quiroga M, Bell GI, Rutter WJ. Isolation and nucleotide sequence of a cDNA encoding the precursor of mouse nerve growth factor. Nature. 1983 Apr 7;302(5908):538–540. [PubMed]
  • Ip NY, Yancopoulos GD. Neurotrophic factors and their receptors. Ann Neurol. 1994;35 (Suppl):S13–S16. [PubMed]
  • Barbacid M. Neurotrophic factors and their receptors. Curr Opin Cell Biol. 1995 Apr;7(2):148–155. [PubMed]
  • Bresnahan PA, Leduc R, Thomas L, Thorner J, Gibson HL, Brake AJ, Barr PJ, Thomas G. Human fur gene encodes a yeast KEX2-like endoprotease that cleaves pro-beta-NGF in vivo. J Cell Biol. 1990 Dec;111(6 Pt 2):2851–2859. [PMC free article] [PubMed]
  • Lin LF, Doherty DH, Lile JD, Bektesh S, Collins F. GDNF: a glial cell line-derived neurotrophic factor for midbrain dopaminergic neurons. Science. 1993 May 21;260(5111):1130–1132. [PubMed]
  • Seidah NG, Chrétien M, Day R. The family of subtilisin/kexin like pro-protein and pro-hormone convertases: divergent or shared functions. Biochimie. 1994;76(3-4):197–209. [PubMed]
  • Steiner DF, Smeekens SP, Ohagi S, Chan SJ. The new enzymology of precursor processing endoproteases. J Biol Chem. 1992 Nov 25;267(33):23435–23438. [PubMed]
  • Van de Ven WJ, Roebroek AJ, Van Duijnhoven HL. Structure and function of eukaryotic proprotein processing enzymes of the subtilisin family of serine proteases. Crit Rev Oncog. 1993;4(2):115–136. [PubMed]
  • Nakagawa T, Murakami K, Nakayama K. Identification of an isoform with an extremely large Cys-rich region of PC6, a Kex2-like processing endoprotease. FEBS Lett. 1993 Jul 26;327(2):165–171. [PubMed]
  • Lusson J, Vieau D, Hamelin J, Day R, Chrétien M, Seidah NG. cDNA structure of the mouse and rat subtilisin/kexin-like PC5: a candidate proprotein convertase expressed in endocrine and nonendocrine cells. Proc Natl Acad Sci U S A. 1993 Jul 15;90(14):6691–6695. [PMC free article] [PubMed]
  • Day R, Schafer MK, Cullinan WE, Watson SJ, Chrétien M, Seidah NG. Region specific expression of furin mRNA in the rat brain. Neurosci Lett. 1993 Jan 4;149(1):27–30. [PubMed]
  • Schäfer MK, Day R, Cullinan WE, Chrétien M, Seidah NG, Watson SJ. Gene expression of prohormone and proprotein convertases in the rat CNS: a comparative in situ hybridization analysis. J Neurosci. 1993 Mar;13(3):1258–1279. [PubMed]
  • Dong W, Marcinkiewicz M, Vieau D, Chrétien M, Seidah NG, Day R. Distinct mRNA expression of the highly homologous convertases PC5 and PACE4 in the rat brain and pituitary. J Neurosci. 1995 Mar;15(3 Pt 1):1778–1796. [PubMed]
  • Day R, Schafer MK, Watson SJ, Chrétien M, Seidah NG. Distribution and regulation of the prohormone convertases PC1 and PC2 in the rat pituitary. Mol Endocrinol. 1992 Mar;6(3):485–497. [PubMed]
  • Seidah NG, Day R, Hamelin J, Gaspar A, Collard MW, Chrétien M. Testicular expression of PC4 in the rat: molecular diversity of a novel germ cell-specific Kex2/subtilisin-like proprotein convertase. Mol Endocrinol. 1992 Oct;6(10):1559–1570. [PubMed]
  • Molloy SS, Thomas L, VanSlyke JK, Stenberg PE, Thomas G. Intracellular trafficking and activation of the furin proprotein convertase: localization to the TGN and recycling from the cell surface. EMBO J. 1994 Jan 1;13(1):18–33. [PMC free article] [PubMed]
  • Malide D, Seidah NG, Chrétien M, Bendayan M. Electron microscopic immunocytochemical evidence for the involvement of the convertases PC1 and PC2 in the processing of proinsulin in pancreatic beta-cells. J Histochem Cytochem. 1995 Jan;43(1):11–19. [PubMed]
  • Hosaka M, Nagahama M, Kim WS, Watanabe T, Hatsuzawa K, Ikemizu J, Murakami K, Nakayama K. Arg-X-Lys/Arg-Arg motif as a signal for precursor cleavage catalyzed by furin within the constitutive secretory pathway. J Biol Chem. 1991 Jul 5;266(19):12127–12130. [PubMed]
  • Seidah NG, Chrétien M. Pro-protein convertases of subtilisin/kexin family. Methods Enzymol. 1994;244:175–188. [PubMed]
  • Edwards RH, Selby MJ, Mobley WC, Weinrich SL, Hruby DE, Rutter WJ. Processing and secretion of nerve growth factor: expression in mammalian cells with a vaccinia virus vector. Mol Cell Biol. 1988 Jun;8(6):2456–2464. [PMC free article] [PubMed]
  • Benjannet S, Rondeau N, Day R, Chrétien M, Seidah NG. PC1 and PC2 are proprotein convertases capable of cleaving proopiomelanocortin at distinct pairs of basic residues. Proc Natl Acad Sci U S A. 1991 May 1;88(9):3564–3568. [PMC free article] [PubMed]
  • Benjannet S, Rondeau N, Paquet L, Boudreault A, Lazure C, Chrétien M, Seidah NG. 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. Biochem J. 1993 Sep 15;294(Pt 3):735–743. [PMC free article] [PubMed]
  • Kiefer MC, Tucker JE, Joh R, Landsberg KE, Saltman D, Barr PJ. Identification of a second human subtilisin-like protease gene in the fes/fps region of chromosome 15. DNA Cell Biol. 1991 Dec;10(10):757–769. [PubMed]
  • Benjannet S, Reudelhuber T, Mercure C, Rondeau N, Chrétien M, Seidah NG. 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. J Biol Chem. 1992 Jun 5;267(16):11417–11423. [PubMed]
  • Helman LJ, Ahn TG, Levine MA, Allison A, Cohen PS, Cooper MJ, Cohn DV, Israel MA. Molecular cloning and primary structure of human chromogranin A (secretory protein I) cDNA. J Biol Chem. 1988 Aug 15;263(23):11559–11563. [PubMed]
  • Linard CG, Mbikay M, Seidah NG, Chretien M. Primary structure of mouse chromogranin B deduced from cDNA sequence. Nucleic Acids Res. 1990 Mar 11;18(5):1298–1298. [PMC free article] [PubMed]
  • Hoflehner J, Eder U, Laslop A, Seidah NG, Fischer-Colbrie R, Winkler H. Processing of secretogranin II by prohormone convertases: importance of PC1 in generation of secretoneurin. FEBS Lett. 1995 Mar 6;360(3):294–298. [PubMed]
  • Takahashi S, Kasai K, Hatsuzawa K, Kitamura N, Misumi Y, Ikehara Y, Murakami K, Nakayama K. A mutation of furin causes the lack of precursor-processing activity in human colon carcinoma LoVo cells. Biochem Biophys Res Commun. 1993 Sep 15;195(2):1019–1026. [PubMed]
  • Galanopoulou AS, Seidah NG, Patel YC. Heterologous processing of rat prosomatostatin to somatostatin-14 by PC2: requirement for secretory cell but not the secretion granule. Biochem J. 1995 Oct 1;311(Pt 1):111–118. [PMC free article] [PubMed]
  • Huttner WB, Gerdes HH, Rosa P. The granin (chromogranin/secretogranin) family. Trends Biochem Sci. 1991 Jan;16(1):27–30. [PubMed]
  • Baeuerle PA, Huttner WB. Tyrosine sulfation is a trans-Golgi-specific protein modification. J Cell Biol. 1987 Dec;105(6 Pt 1):2655–2664. [PMC free article] [PubMed]
  • Rosenquist GL, Nicholas HB., Jr Analysis of sequence requirements for protein tyrosine sulfation. Protein Sci. 1993 Feb;2(2):215–222. [PMC free article] [PubMed]
  • Nishi R. Neurotrophic factors: two are better than one. Science. 1994 Aug 19;265(5175):1052–1053. [PubMed]
  • Zheng M, Streck RD, Scott RE, Seidah NG, Pintar JE. The developmental expression in rat of proteases furin, PC1, PC2, and carboxypeptidase E: implications for early maturation of proteolytic processing capacity. J Neurosci. 1994 Aug;14(8):4656–4673. [PubMed]
  • Mbikay M, Seidah NG, Chrétien M, Simpson EM. Chromosomal assignment of the genes for proprotein convertases PC4, PC5, and PACE 4 in mouse and human. Genomics. 1995 Mar 1;26(1):123–129. [PubMed]
  • Varon S, Nomura J, Shooter EM. The isolation of the mouse nerve growth factor protein in a high molecular weight form. Biochemistry. 1967 Jul;6(7):2202–2209. [PubMed]
  • Silverman RE, Bradshaw RA. Nerve growth factor: subunit interactions in the mouse submaxillary gland 7S complex. J Neurosci Res. 1982;8(2-3):127–136. [PubMed]
  • Edwards RH, Selby MJ, Garcia PD, Rutter WJ. Processing of the native nerve growth factor precursor to form biologically active nerve growth factor. J Biol Chem. 1988 May 15;263(14):6810–6815. [PubMed]
  • Murphy RA, Landis SC, Bernanke J, Siminoski K. Absence of the alpha and gamma subunits of 7S nerve growth factor in denervated rodent iris: immunocytochemical studies. Dev Biol. 1986 Apr;114(2):369–380. [PubMed]
  • Mobley WC, Schenker A, Shooter EM. Characterization and isolation of proteolytically modified nerve growth factor. Biochemistry. 1976 Dec 14;15(25):5543–5552. [PubMed]
  • Kornfeld R, Kornfeld S. Assembly of asparagine-linked oligosaccharides. Annu Rev Biochem. 1985;54:631–664. [PubMed]
  • Murphy RA, Chlumecky V, Smillie LB, Carpenter M, Nattriss M, Anderson JK, Rhodes JA, Barker PA, Siminoski K, Campenot RB, et al. Isolation and characterization of a glycosylated form of beta nerve growth factor in mouse submandibular glands. J Biol Chem. 1989 Jul 25;264(21):12502–12509. [PubMed]
  • Winkler H, Fischer-Colbrie R. The chromogranins A and B: the first 25 years and future perspectives. Neuroscience. 1992 Aug;49(3):497–528. [PubMed]
  • Fischer-Colbrie R, Hagn C, Kilpatrick L, Winkler H. Chromogranin C: a third component of the acidic proteins in chromaffin granules. J Neurochem. 1986 Jul;47(1):318–321. [PubMed]

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