Design and synthesis of truncated EGF-A peptides that restore LDL-R recycling in the presence of PCSK9 in vitro

Christina I Schroeder; Joakim E Swedberg; Jane M Withka; K Johan Rosengren; Muharrem Akcan; Daniel J Clayton; Norelle L Daly; Olivier Cheneval; Kris A Borzilleri; Matt Griffor; Ingrid Stock; Barbara Colless; Phillip Walsh; Philip Sunderland; Allan Reyes; Robert Dullea; Mark Ammirati; Shenping Liu; Kim F McClure; Meihua Tu; Samit K Bhattacharya; Spiros Liras; David A Price; David J Craik

doi:10.1016/j.chembiol.2013.11.014

Design and synthesis of truncated EGF-A peptides that restore LDL-R recycling in the presence of PCSK9 in vitro

Chem Biol. 2014 Feb 20;21(2):284-94. doi: 10.1016/j.chembiol.2013.11.014. Epub 2014 Jan 16.

Authors

Christina I Schroeder¹, Joakim E Swedberg¹, Jane M Withka², K Johan Rosengren³, Muharrem Akcan¹, Daniel J Clayton¹, Norelle L Daly¹, Olivier Cheneval¹, Kris A Borzilleri², Matt Griffor², Ingrid Stock⁴, Barbara Colless¹, Phillip Walsh¹, Philip Sunderland¹, Allan Reyes⁵, Robert Dullea⁵, Mark Ammirati², Shenping Liu², Kim F McClure⁵, Meihua Tu⁵, Samit K Bhattacharya⁵, Spiros Liras⁵, David A Price⁵, David J Craik⁶

Affiliations

¹ Institute for Molecular Bioscience, The University of Queensland, Brisbane, 4072 QLD, Australia.
² Structural Biology and Biophysics, Pfizer Global Research and Development, Eastern Point Road, Groton, CT 06340, USA.
³ School of Biomedical Sciences, The University of Queensland, Brisbane, 4072 QLD, Australia.
⁴ PDM-NCE, Pfizer Global Research and Development, Eastern Point Road, Groton, CT 06340, USA.
⁵ CVMED, Pfizer Global Research and Development, 620 Memorial Drive, Cambridge, MA 02139, USA.
⁶ Institute for Molecular Bioscience, The University of Queensland, Brisbane, 4072 QLD, Australia. Electronic address: [email protected].

PMID: 24440079
DOI: 10.1016/j.chembiol.2013.11.014

Abstract

Disrupting the binding interaction between proprotein convertase (PCSK9) and the epidermal growth factor-like domain A (EGF-A domain) in the low-density lipoprotein receptor (LDL-R) is a promising strategy to promote LDL-R recycling and thereby lower circulating cholesterol levels. In this study, truncated 26 amino acid EGF-A analogs were designed and synthesized, and their structures were analyzed in solution and in complex with PCSK9. The most potent peptide had an increased binding affinity for PCSK9 (KD = 0.6 μM) compared with wild-type EGF-A (KD = 1.2 μM), and the ability to increase LDL-R recycling in the presence of PCSK9 in a cell-based assay.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Binding Sites
Cell Line
Cholesterol / metabolism
Epidermal Growth Factor / chemistry
Fluorescence Resonance Energy Transfer
Humans
Molecular Dynamics Simulation
Molecular Sequence Data
Mutagenesis
Peptides / chemical synthesis
Peptides / chemistry
Peptides / metabolism*
Proprotein Convertase 9
Proprotein Convertases / chemistry
Proprotein Convertases / genetics
Proprotein Convertases / metabolism*
Protein Binding
Protein Structure, Secondary
Protein Structure, Tertiary
Receptors, LDL / metabolism*
Serine Endopeptidases / chemistry
Serine Endopeptidases / genetics
Serine Endopeptidases / metabolism*

Substances

Peptides
Receptors, LDL
Epidermal Growth Factor
Cholesterol
PCSK9 protein, human
Proprotein Convertase 9
Proprotein Convertases
Serine Endopeptidases