PDBsum entry 5hhc

De novo protein

PDB id: 5hhc

Contents

Protein chains

95 a.a.

66 a.a.

Ligands

GOL ×2

Waters ×177

PDB id:

5hhc

Name:

De novo protein

Title:

Crystal structure of chemically synthesized heterochiral {rfx037 plus vegf-a} protein complex in space group p21/n

Structure:

Vascular endothelial growth factor a. Chain: a, b. Synonym: vegf-a,vascular permeability factor,vpf. Engineered: yes. D- vascular endothelial growth factor-a. Chain: c, d. Engineered: yes

Source:

Synthetic: yes. Homo sapiens. Human. Organism_taxid: 9606. Other_details: chemical synthesis. Synthetic construct. Organism_taxid: 32630

Resolution:

2.10Å R-factor: 0.263 R-free: 0.307

Authors:

M.Uppalapati,D.J.Lee,K.Mandal,S.B.H.Kent,S.Sidhu

Key ref:

M.Uppalapati et al. (2016). A Potent d-Protein Antagonist of VEGF-A is Nonimmunogenic, Metabolically Stable, and Longer-Circulating in Vivo. Acs Chem Biol, 11, 1058-1065. PubMed id: 26745345 DOI: 10.1021/acschembio.5b01006

Date:

10-Jan-16 Release date: 09-Mar-16

Protein chains

P15692  (VEGFA_HUMAN) -  Vascular endothelial growth factor A, long form from Homo sapiens

Seq: 395 a.a.

Struc: 95 a.a.

Protein chains

No UniProt id for this chain

Struc: 66 a.a.

DOI no: 10.1021/acschembio.5b01006

Acs Chem Biol 11:1058-1065 (2016)

PubMed id: 26745345

A Potent d-Protein Antagonist of VEGF-A is Nonimmunogenic, Metabolically Stable, and Longer-Circulating in Vivo.

M.Uppalapati, D.J.Lee, K.Mandal, H.Li, L.P.Miranda, J.Lowitz, J.Kenney, J.J.Adams, D.Ault-Riché, S.B.Kent, S.S.Sidhu.

ABSTRACT

Polypeptides composed entirely of d-amino acids and the achiral amino acid glycine (d-proteins) inherently have in vivo properties that are proposed to be near-optimal for a large molecule therapeutic agent. Specifically, d-proteins are resistant to degradation by proteases and are anticipated to be nonimmunogenic. Furthermore, d-proteins are manufactured chemically and can be engineered to have other desirable properties, such as improved stability, affinity, and pharmacokinetics. Thus, a well-designed d-protein therapeutic would likely have significant advantages over l-protein drugs. Toward the goal of developing d-protein therapeutics, we previously generated RFX001.D, a d-protein antagonist of natural vascular endothelial growth factor A (VEGF-A) that inhibited binding to its receptor. However, RFX001.D is unstable at physiological temperatures (Tm = 33 °C). Here, we describe RFX037.D, a variant of RFX001.D with extreme thermal stability (Tm > 95 °C), high affinity for VEGF-A (Kd = 6 nM), and improved receptor blocking. Comparison of the two enantiomeric forms of RFX037 revealed that the d-protein is more stable in mouse, monkey, and human plasma and has a longer half-life in vivo in mice. Significantly, RFX037.D was nonimmunogenic in mice, whereas the l-enantiomer generated a strong immune response. These results confirm the potential utility of synthetic d-proteins as alternatives to therapeutic antibodies.