Point mutations at the catalytic site of PCSK9 inhibit folding, autoprocessing, and interaction with the LDL receptor
Document Type
Article
Publication Date
8-18-2016
Publication Source
Protein Science
Abstract
Circulating low-density lipoprotein cholesterol (LDLc) is regulated by membrane-bound LDL receptor (LDLr). Upon LDLc and LDLr interaction the complex is internalized by the cell, leading to LDLc degradation and LDLr recycling back to the cell surface. The proprotein convertase subtilisin/ kexin type 9 (PCSK9) protein regulates this cycling. PCSK9 is secreted from the cell and binds LDLr. When the complex is internalized, PCSK9 prevents LDLr from shuttling back to the surface and instead targets it for degradation. PCSK9 is a serine protease expressed as a zymogen that undergoes autoproteolysis, though the two resulting protein domains remain stably associated as a heterodimer. This PCSK9 autoprocessing is required for the protein to be secreted from the cell. To date, direct analysis of PCSK9 autoprocessing has proven challenging, as no catalytically active zymogen has been isolated. A PCSK9 loss-of-function point mutation (Q152H) that reduces LDLc levels two-fold was identified in a patient population. LDLc reduction was attributed to a lack of PCSK9(Q152H) autoprocessing preventing secretion of the protein. We have isolated a zymogen form of PCSK9, PCSK9(Q152H), and a related mutation (Q152N), that can undergo slow autoproteolysis. We show that the point mutation prevents the formation of the mature form of PCSK9 by hindering folding, reducing the rate of autoproteolysis, and destabilizing the heterodimeric form of the protein. In addition, we show that the zymogen form of PCSK9 adopts a structure that is distinct from the processed form and is unable to bind a mimetic peptide based on the EGF-A domain of the LDLr.
Inclusive pages
2018-2027
ISBN/ISSN
0961-8368
Copyright
Copyright © 2016, Protein Society
Publisher
John Wiley & Sons
Volume
25
Peer Reviewed
yes
Issue
11
Keywords
PCSK9, autoprocessing, autoproteolysis, zymogen, point mutations, serine protease, low-density lipoprotein receptor, LDLR, protein refolding
eCommons Citation
Garvie, Colin W.; Fraley, Cara V.; Elowe, Nadine H.; Culyba, Elizabeth K.; Lemke, Christopher T.; Hubbard, Brian K.; Kaushik, Virendar K.; and Daniels, Douglas S., "Point mutations at the catalytic site of PCSK9 inhibit folding, autoprocessing, and interaction with the LDL receptor" (2016). Chemistry Faculty Publications. 98.
https://ecommons.udayton.edu/chm_fac_pub/98
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