Subcellular localization and partial purification of prelamin A endoprotease: an enzyme which catalyzes the conversion of farnesylated prelamin A to mature lamin A.
The nuclear lamina protein, lamin A is produced by proteolytic cleavage of a 74 kDa precursor protein, prelamin A. The conversion of this precursor to mature lamin A is mediated by a specific endoprotease, prelamin A endoprotease. Subnuclear fractionation indicates that the prelamin A endoprotease is localized at the nuclear membrane. The enzyme appears to be an integral membrane protein, as it can only be removed from the nuclear envelope with detergent. It is effectively solubilized by the detergent n-octyl-beta-D-glucopyranoside and can be partially-purified (approximately 1200-fold) by size exclusion and cation exchange (Mono S) chromatography. Prelamin A endoprotease from HeLa cells was eluted from Mono S with 0.3 M sodium chloride as a single peak of activity. SDS-PAGE analysis of this prelamin A endoprotease preparation shows that it contains one major polypeptide at 65 kDa and smaller amounts of a second 68 kDa polypeptide. Inhibition of the enzyme activity in this preparation by specific serine protease inhibitors is consistent with the enzyme being a serine protease.
Kilic
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Department of Biochemistry and Molecular Biology, East Tennessee State University, James H. Quillen College of Medicine, Johnson City 37614, USA. fusun.kilic@yale.edu
Johnson
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DA
Sinensky
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M
D000180:Acrylic Resins; D003902:Detergents; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000302:isolation & purification*; D005960:Glucosides / Q000378:metabolism; D006367:HeLa Cells / Q000201:enzymology*; D006801:Humans; D034904:Lamin Type A; D034882:Lamins; D009687:Nuclear Proteins / Q000378:metabolism*; D011498:Protein Precursors / Q000378:metabolism*; D017368:Protein Prenylation; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology; D013347:Subcellular Fractions / Q000201:enzymology
D000180:Acrylic Resins; D003902:Detergents; D005960:Glucosides; D034904:Lamin Type A; D034882:Lamins; D009687:Nuclear Proteins; D011498:Protein Precursors; D015842:Serine Proteinase Inhibitors; C090764:prelamin A; C018619:octyl-beta-D-glucoside; C023049:sephacryl S 200; D010450:Endopeptidases; C104823:prelamin A endoprotease