PDBsum entry 4ov6

Hydrolase/protein binding

PDB id: 4ov6

Contents

Protein chains

92 a.a.

281 a.a.

99 a.a.

Ligands

EDO ×5

PG4 ×2

Waters ×130

PDB id:

4ov6

Name:

Hydrolase/protein binding

Title:

Crystal structure of pcsk9(53-451) with adnectin

Structure:

Proprotein convertase subtilisin/kexin type 9. Chain: a, d. Fragment: prodomain (unp residues 60-152). Synonym: pcsk9, neural apoptosis-regulated convertase 1, narc-1, proprotein convertase 9, pc9, subtilisin/kexin-like protease pc9. Engineered: yes. Proprotein convertase subtilisin/kexin type 9. Chain: b, e. Fragment: catalytic domain (unp residues 153-446).

Source:

Homo sapiens. Human. Organism_taxid: 9606. Gene: pcsk9, narc1, psec0052. Expressed in: trichoplusia ni. Expression_system_taxid: 7111. Expression_system_cell_line: high five cells. Expressed in: escherichia coli. Expression_system_taxid: 469008.

Resolution:

2.69Å R-factor: 0.201 R-free: 0.231

Authors:

J.A.Khan

Key ref:

T.Mitchell et al. (2014). Pharmacologic profile of the Adnectin BMS-962476, a small protein biologic alternative to PCSK9 antibodies for low-density lipoprotein lowering. J Pharmacol Exp Ther, 350, 412-424. PubMed id: 24917546 DOI: 10.1124/jpet.114.214221

Date:

20-Feb-14 Release date: 02-Jul-14

Protein chains

Q8NBP7  (PCSK9_HUMAN) -  Proprotein convertase subtilisin/kexin type 9 from Homo sapiens

Seq: 692 a.a.

Struc: 92 a.a.

Protein chains

Q8NBP7  (PCSK9_HUMAN) -  Proprotein convertase subtilisin/kexin type 9 from Homo sapiens

Seq: 692 a.a.

Struc: 281 a.a.

Protein chains

A0A075B5G3  (A0A075B5G3_HUMAN) -  Adnectin from Homo sapiens

Seq: 99 a.a.

Struc: 99 a.a.

Enzyme reactions

• Enzyme class: Chains A, B, D, E: E.C.3.4.21.- - ?????

DOI no: 10.1124/jpet.114.214221

J Pharmacol Exp Ther 350:412-424 (2014)

PubMed id: 24917546

Pharmacologic profile of the Adnectin BMS-962476, a small protein biologic alternative to PCSK9 antibodies for low-density lipoprotein lowering.

T.Mitchell, G.Chao, D.Sitkoff, F.Lo, H.Monshizadegan, D.Meyers, S.Low, K.Russo, R.DiBella, F.Denhez, M.Gao, J.Myers, G.Duke, M.Witmer, B.Miao, S.P.Ho, J.Khan, R.A.Parker.

ABSTRACT

Proprotein convertase subtilisin kexin-9 (PCSK9) is an important pharmacological target for decreasing low-density lipoprotein (LDL) in cardiovascular disease, although seemingly inaccessible to small molecule approaches. Compared with therapeutic IgG antibodies currently in development, targeting circulating PCSK9 with smaller molecular scaffolds could offer different profiles and reduced dose burdens. This inspired genesis of PCSK9-binding Adnectins, a protein family derived from human fibronectin-10th-type III-domain and engineered for high-affinity target binding. BMS-962476, an ∼11-kDa polypeptide conjugated to polyethylene glycol to enhance pharmacokinetics, binds with subnanomolar affinity to human. The X-ray cocrystal structure of PCSK9 with a progenitor Adnectin shows ∼910 Å(2) of PCSK9 surface covered next to the LDL receptor binding site, largely by residues of a single loop of the Adnectin. In hypercholesterolemic, overexpressing human PCSK9 transgenic mice, BMS-962476 rapidly lowered cholesterol and free PCSK9 levels. In genomic transgenic mice, BMS-962476 potently reduced free human PCSK9 (ED50 ∼0.01 mg/kg) followed by ∼2-fold increases in total PCSK9 before return to baseline. Treatment of cynomolgus monkeys with BMS-962476 rapidly suppressed free PCSK9 >99% and LDL-cholesterol ∼55% with subsequent 6-fold increase in total PCSK9, suggesting reduced clearance of circulating complex. Liver sterol response genes were consequently downregulated, following which LDL and total PCSK9 returned to baseline. These studies highlight the rapid dynamics of PCSK9 control over LDL and liver cholesterol metabolism and characterize BMS-962476 as a potent and efficacious PCSK9 inhibitor.