6DRF

Structure of human Retinal Degeneration 3(RD3) Protein

6DRF の概要

• エントリーDOI: 10.2210/pdb6drf/pdb
• NMR情報: BMRB: 27305
• 分子名称: Protein RD3 (1 entity in total)
• 機能のキーワード: sensory transduction, vision, signaling protein
• 由来する生物種: Homo sapiens (Human)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 16601.79

構造登録者

Yu, Q.,Lim, S.,Peshenko, I.,Cudia, D.,Dizhoor, A.M.,Ames, J.B. (登録日: 2018-06-11, 公開日: 2019-02-06, 最終更新日: 2024-05-15)

主引用文献

Peshenko, I.V.,Yu, Q.,Lim, S.,Cudia, D.,Dizhoor, A.M.,Ames, J.B.
Retinal degeneration 3 (RD3) protein, a retinal guanylyl cyclase regulator, forms a monomeric and elongated four-helix bundle.
J. Biol. Chem., 294:2318-2328, 2019

PubMed Abstract: Retinal degeneration 3 (RD3) protein promotes accumulation of retinal membrane guanylyl cyclase (RetGC) in the photoreceptor outer segment and suppresses RetGC activation by guanylyl cyclase-activating proteins (GCAPs). Mutations truncating RD3 cause severe congenital blindness by preventing the inhibitory binding of RD3 to the cyclase. The high propensity of RD3 to aggregate in solution has prevented structural analysis. Here, we produced a highly soluble variant of human RD3 (residues 18-160) that is monomeric and can still bind and negatively regulate RetGC. The NMR solution structure of RD3 revealed an elongated backbone structure (70 Å long and 30 Å wide) consisting of a four-helix bundle with a long unstructured loop between helices 1 and 2. The structure reveals that RD3 residues previously implicated in the RetGC binding map to a localized and contiguous area on the structure, involving a loop between helices 2 and 3 and adjacent parts of helices 3 and 4. The NMR structure of RD3 was validated by mutagenesis. Introducing Trp or Phe to replace Cys or Leu, respectively, disrupts packing in the hydrophobic core and lowers RD3's apparent affinity for RetGC1. Introducing a positive charge at the interface (Glu to Lys) also lowered the affinity. Conversely, introducing Val in place of Cys stabilized the hydrophobic core and increased the RD3 affinity for the cyclase. The NMR structure of RD3 presented here provides a structural basis for elucidating RD3-RetGC interactions relevant for normal vision or blindness.

PubMed: 30559291
DOI: 10.1074/jbc.RA118.006106

実験手法

SOLUTION NMR