Prelamin A endoproteolytic processing in vitro by recombinant Zmpste24

Douglas P. Corrigan, Danuta Kuszczak, Antonio E. Rusinol, Douglas P. Thewke, Christine A. Hrycyna, Susan Michaelis, Michael S. Sinensky

Research output: Contribution to journalArticle


The nuclear lamins form a karyoskeleton providing structural rigidity to the nucleus. One member of the lamin family, lamin A, is first synthesized as a 74 kDa precursor, prelamin A. After the endopeptidase and methylation reactions which occur after farnesylation of the CAAX-box cysteine, there is a second endoproteolysis that occurs 15 amino acids upstream from the C-terminal farnesylated cysteine residue. Studies with knockout mice have implicated the enzyme Zmpste24 (Face-1) as a suitable candidate to perform one or both of these proteolytic reactions. Evidence has been presented elsewhere establishing that Zmpste24 possesses a zinc-dependent CAAX endopeptidase activity. In the present study, we confirm this CAAX endopeptidase activity with recombinant, membrane-reconstituted Zmpste24 and show that it can accept a prelamin A farnesylated tetrapeptide as substrate. To monitor the second upstream endoproteolytic cleavage of prelamin A, we expressed a 33 kDa prelamin A C-terminal tail in insect cells. We demonstrate that this purified substrate possesses a C-terminal farnesylated and carboxyl-methylated cysteine and, therefore, constitutes a valid substrate for assaying the second endoproteolytic step in lamin A maturation. With this substrate, we demonstrate that insect cell membranes bearing recombinant Zmpste24 can also catalyse the second upstream endoproteolytic cleavage.

Original languageEnglish (US)
Pages (from-to)129-138
Number of pages10
JournalBiochemical Journal
Issue number1
StatePublished - Apr 1 2005


  • Endoproteolysis
  • Face-1
  • Lamin A
  • Prelamin A
  • Prenylation
  • Zmpste24

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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  • Cite this

    Corrigan, D. P., Kuszczak, D., Rusinol, A. E., Thewke, D. P., Hrycyna, C. A., Michaelis, S., & Sinensky, M. S. (2005). Prelamin A endoproteolytic processing in vitro by recombinant Zmpste24. Biochemical Journal, 387(1), 129-138.