6OU3

Crystal Structure of the D478S Variant of the Myocilin Olfactomedin Domain

6OU3 の概要

• エントリーDOI: 10.2210/pdb6ou3/pdb
• 分子名称: Myocilin, SODIUM ION, GLYCEROL, ... (4 entities in total)
• 機能のキーワード: olfactomedin myocilin beta propeller, protein binding
• 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: 31376.15

構造登録者

Hill, S.E.,Kwon, M.S.,Lieberman, R.L. (登録日: 2019-05-03, 公開日: 2019-07-03, 最終更新日: 2024-11-06)

主引用文献

Hill, S.E.,Kwon, M.S.,Martin, M.D.,Suntharalingam, A.,Hazel, A.,Dickey, C.A.,Gumbart, J.C.,Lieberman, R.L.
Stable calcium-free myocilin olfactomedin domain variants reveal challenges in differentiating between benign and glaucoma-causing mutations.
J.Biol.Chem., 294:12717-12728, 2019

PubMed Abstract: Nonsynonymous gene mutations can be beneficial, neutral, or detrimental to the stability, structure, and biological function of the encoded protein, but the effects of these mutations are often not readily predictable. For example, the β-propeller olfactomedin domain of myocilin (mOLF) exhibits a complex interrelationship among structure(s), stability, and aggregation. Numerous mutations within mOLF are linked to glaucoma; the resulting variants are less stable, aggregation-prone, and sequestered intracellularly, causing cytotoxicity. Here, we report the first stable mOLF variants carrying substitutions in the calcium-binding site that exhibit solution characteristics indistinguishable from those of glaucoma variants. Crystal structures of these stable variants at 1.8-2.0-Å resolution revealed features that we could not predict by molecular dynamics simulations, including loss of loop structure, helix unwinding, and a blade shift. Double mutants that combined a stabilizing substitution and a selected glaucoma-causing single-point mutant rescued folding and stability defects. In the context of full-length myocilin, secretion of stable single variants was indistinguishable from that of the WT protein, and the double mutants were secreted to varying extents. In summary, our finding that mOLF can tolerate particular substitutions that render the protein stable despite a conformational switch emphasizes the complexities in differentiating between benign and glaucoma-causing variants and provides new insight into the possible biological function of myocilin.

PubMed: 31270212
DOI: 10.1074/jbc.RA119.009419

実験手法

X-RAY DIFFRACTION (1.796 Å)