Characterisation of de novo mutations in the C-terminal domain of proprotein convertase subtilisin/kexin type 9

Protein Engineering, Design & Selection : PEDS
Stefan GeschwindnerWolfgang Knecht

Abstract

Proprotein convertase subtilisin/kexin type 9 (PCSK9) promotes the degradation of the hepatic low-density lipoprotein receptor (LDL-R) and is therefore a prominent therapeutic target for reducing LDL-cholesterol. The C-terminal domain of PCSK9 is unlikely to be involved in a direct extracellular interaction with the LDL-R. We probed the importance of the C-terminus for the degradation of the LDL-R by designing seven de novo mutants of PCSK9 that fill potential druggable cavities. The mutants were tested for their ability to diminish LDL uptake in human HepG2 cells and for affinity towards a calcium independent mutant of the EGF(A) domain of the human LDL-R. The later was done by a newly developed surface plasmon resonance-based assay format. We identified three mutant proteins (G517R, V610R and V644R) with decreased ability to block LDL uptake into HepG2 cells. These mutations define areas outside the direct interaction area between PCSK9 and the LDL-R that could be targeted to inhibit the PCSK9 triggered degradation of the LDL-R. We also describe the mechanistic rationalisation of the affinity changes seen with the natural occurring human D374Y (gain of function) mutation causing severe hypercholesterolaemia. The action of thi...Continue Reading

References

Jul 5, 2002·Journal of Molecular Biology·Matthew P JacobsonBarry Honig
Jan 29, 2003·Proceedings of the National Academy of Sciences of the United States of America·Nabil G SeidahMichel Chretien
May 6, 2003·Nature Genetics·Marianne AbifadelCatherine Boileau
Sep 27, 2003·Proceedings of the National Academy of Sciences of the United States of America·Jay D HortonJoseph L Goldstein
Apr 3, 2004·Nature Reviews. Drug Discovery·Michelle R Arkin, James A Wells
May 1, 2004·Proceedings of the National Academy of Sciences of the United States of America·Kara N Maxwell, Jan L Breslow
Apr 2, 2005·Journal of Medicinal Chemistry·Philip J HajdukStephen W Fesik
Feb 9, 2006·American Journal of Human Genetics·Ingrid K KotowskiHelen H Hobbs
Mar 24, 2006·The New England Journal of Medicine·Jonathan C CohenHelen H Hobbs
Jan 12, 2007·Trends in Biochemical Sciences·Jay D HortonHelen H Hobbs
Mar 27, 2007·Chemical Biology & Drug Design·Tom Halgren
Apr 17, 2007·Nature Structural & Molecular Biology·David CunninghamXiayang Qiu
May 16, 2007·Structure·Derek E PiperNigel P C Walker
Jul 17, 2007·PLoS Computational Biology·Yanay Ofran, Burkhard Rost
Sep 7, 2007·Proceedings of the National Academy of Sciences of the United States of America·Eric N HamptonGlen Spraggon
Feb 6, 2008·Proceedings of the National Academy of Sciences of the United States of America·Hyock Joo KwonJohann Deisenhofer
Mar 21, 2008·Journal of Lipid Research·Shilpa PanditTimothy S Fisher
Jul 31, 2008·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·Ahmed ZaidAnnik Prat
Sep 19, 2008·The Journal of Biological Chemistry·Gaétan MayerNabil G Seidah
Mar 21, 2009·Arteriosclerosis, Thrombosis, and Vascular Biology·Joseph L Goldstein, Michael S Brown
Jun 3, 2009·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·Patrick LabontéNabil G Seidah
May 11, 2010·Journal of Lipid Research·Mali LiuCelina V Zerbinatti
Feb 18, 2011·Biochemical and Biophysical Research Communications·Øystein L HollaTrond P Leren
Jul 21, 2011·Journal of Lipid Research·Øystein L HollaTrond P Leren
Nov 15, 2011·Biochemical and Biophysical Research Communications·Yanqun ChenJian Wang
Jun 2, 2012·Current Pharmaceutical Design·Justin F Bower, Andrew Pannifer
Jun 12, 2012·Nature Reviews. Drug Discovery·Nabil G Seidah, Annik Prat
Jun 26, 2012·Journal of Molecular Biology·Yingnan ZhangDaniel Kirchhofer
Jul 26, 2012·Molecular Therapy : the Journal of the American Society of Gene Therapy·Belete A DesimmieFrauke Christ
Aug 22, 2012·Current Opinion in Lipidology·David A MaraisGilles Lambert
Nov 21, 2013·Journal of Molecular Biology·Felix SchieleHerbert Nar

❮ Previous
Next ❯

Citations

Jan 18, 2020·The Journal of Biological Chemistry·Samantha K SarkarThomas A Lagace
Nov 12, 2020·Frontiers in Genetics·Qianyun GuoYujie Zhou

❮ Previous
Next ❯

Related Concepts

Related Feeds

ApoE, Lipids & Cholesterol

Serum cholesterol, triglycerides, apolipoprotein B (APOB)-containing lipoproteins (very low-density lipoprotein (VLDL), immediate-density lipoprotein (IDL), and low-density lipoprotein (LDL), lipoprotein A (LPA)) and the total cholesterol/high-density lipoprotein (HDL) cholesterol ratio are all connected in diseases. Here is the latest research.