Characterization of PC2, a mammalian Kex2 homologue, following expression of the cDNA in microinjected Xenopus oocytes

K I Shennan, S P Smeekens, D F Steiner, K Docherty

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

A human insulinoma cDNA (PC2) that encodes a protein homologous to the Kex2/subtilisin-like proteinases has recently been described [1990, J. Biol. Chem. 265, 2997-3000]. In order to characterise the associated proteinase activity, mRNA encoding PC2 was synthesised in vitro and microinjected into Xenopus oocytes. The proteinase activity released into the media from oocytes microinjected with PC2 mRNA was assayed using small peptide fluorogenic substrates. Boc.Gln.Arg.Arg aminomethyl coumarin was hydrolysed in a Ca(2+)-dependent manner, but substrate analogues bearing a single basic aminoacid were not. The substrate specificity, inhibitor profile, and pH optimum of 5.5 were compatible with an involvement of PC2 in prohormone processing in mammalian cells.
Original languageEnglish
Pages (from-to)277-80
Number of pages4
JournalFEBS Letters
Volume284
Issue number2
Publication statusPublished - 24 Jun 1991

Fingerprint

Xenopus
Oocytes
Peptide Hydrolases
Complementary DNA
Bearings (structural)
Subtilisin
Messenger RNA
Insulinoma
Substrates
Substrate Specificity
Fluorescent Dyes
Cells
Peptides
Processing
Proteins

Keywords

  • Animals
  • Calcium
  • DNA
  • Enzyme Activation
  • Gene Expression
  • Humans
  • Insulinoma
  • Microinjections
  • Oocytes
  • Pancreatic Neoplasms
  • Proprotein Convertase 2
  • Proprotein Convertases
  • Protein Precursors
  • RNA, Messenger
  • Saccharomyces cerevisiae Proteins
  • Serine Endopeptidases
  • Serine Proteinase Inhibitors
  • Substrate Specificity
  • Subtilisins
  • Transfection
  • Xenopus laevis

Cite this

Characterization of PC2, a mammalian Kex2 homologue, following expression of the cDNA in microinjected Xenopus oocytes. / Shennan, K I; Smeekens, S P; Steiner, D F; Docherty, K.

In: FEBS Letters, Vol. 284, No. 2, 24.06.1991, p. 277-80.

Research output: Contribution to journalArticle

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abstract = "A human insulinoma cDNA (PC2) that encodes a protein homologous to the Kex2/subtilisin-like proteinases has recently been described [1990, J. Biol. Chem. 265, 2997-3000]. In order to characterise the associated proteinase activity, mRNA encoding PC2 was synthesised in vitro and microinjected into Xenopus oocytes. The proteinase activity released into the media from oocytes microinjected with PC2 mRNA was assayed using small peptide fluorogenic substrates. Boc.Gln.Arg.Arg aminomethyl coumarin was hydrolysed in a Ca(2+)-dependent manner, but substrate analogues bearing a single basic aminoacid were not. The substrate specificity, inhibitor profile, and pH optimum of 5.5 were compatible with an involvement of PC2 in prohormone processing in mammalian cells.",
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KW - DNA

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KW - Gene Expression

KW - Humans

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KW - Microinjections

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KW - Pancreatic Neoplasms

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KW - Saccharomyces cerevisiae Proteins

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KW - Subtilisins

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