2WNP

M-ficolin mutant Y271F

2WNP の概要

• エントリーDOI: 10.2210/pdb2wnp/pdb
• 関連するPDBエントリー: 2JHH 2JHI 2JHK 2JHL 2JHM
• 分子名称: FICOLIN-1, CALCIUM ION, ISOPROPYL ALCOHOL, ... (4 entities in total)
• 機能のキーワード: glycoprotein, innate immunity, fibrinogen-like domain, carbohydrate recognition, lectin, immune system
• 由来する生物種: HOMO SAPIENS (HUMAN)
• 細胞内の位置: Secreted: O00602
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 24329.99

構造登録者

Gout, E.,Garlatti, V.,Smith, D.F.,Lacroix, M.,Dumestre-Perard, C.,Lunardi, T.,Arlaud, G.J.,Gaboriaud, C.,Thielens, N.M. (登録日: 2009-07-16, 公開日: 2009-12-22, 最終更新日: 2024-11-13)

主引用文献

Gout, E.,Garlatti, V.,Smith, D.F.,Lacroix, M.,Dumestre-Perard, C.,Lunardi, T.,Martin, L.,Cesbron, J.Y.,Arlaud, G.J.,Gaboriaud, C.,Thielens, N.M.
Carbohydrate Recognition Properties of Human Ficolins: Glycan Array Screening Reveals the Sialic Acid Binding Specificity of M-Ficolin.
J.Biol.Chem., 285:6612-, 2010

PubMed Abstract: Ficolins are oligomeric innate immune recognition proteins consisting of a collagen-like region and a fibrinogen-like recognition domain that bind to pathogen- and apoptotic cell-associated molecular patterns. To investigate their carbohydrate binding specificities, serum-derived L-ficolin and recombinant H- and M-ficolins were fluorescently labeled, and their carbohydrate binding ability was analyzed by glycan array screening. L-ficolin preferentially recognized disulfated N-acetyllactosamine and tri- and tetrasaccharides containing terminal galactose or N-acetylglucosamine. Binding was sensitive to the position and orientation of the bond between N-acetyllactosamine and the adjacent carbohydrate. No significant binding of H-ficolin to any of the 377 glycans probed could be detected, providing further evidence for its poor lectin activity. M-ficolin bound preferentially to 9-O-acetylated 2-6-linked sialic acid derivatives and to various glycans containing sialic acid engaged in a 2-3 linkage. To further investigate the structural basis of sialic acid recognition by M-ficolin, point mutants were produced in which three residues of the fibrinogen domain were replaced by their counterparts in L-ficolin. Mutations G221F and A256V inhibited binding to the 9-O-acetylated sialic acid derivatives, whereas Y271F abolished interaction with all sialic acid-containing glycans. The crystal structure of the Y271F mutant fibrinogen domain was solved, showing that the mutation does not alter the structure of the ligand binding pocket. These analyses reveal novel ficolin ligands such as sulfated N-acetyllactosamine (L-ficolin) and gangliosides (M-ficolin) and provide precise insights into the sialic acid binding specificity of M-ficolin, emphasizing the essential role of Tyr(271) in this respect.

PubMed: 20032467
DOI: 10.1074/JBC.M109.065854

実験手法

X-RAY DIFFRACTION (1.21 Å)