SASDA38: transglutaminase-2 (TGA2) (transglutaminase 2, TGA)

基本情報

• 登録情報: データベース: SASBDB / ID: SASDA38

試料

transglutaminase-2 (TGA2)

• transglutaminase 2 (protein), TGA, Homo sapiens

• 生物種: Homo sapiens (ヒト)
• 引用: ジャーナル: J Biol Chem / 年: 2015; タイトル: Structural Basis for Antigen Recognition by Transglutaminase 2-specific Autoantibodies in Celiac Disease.; 著者: Xi Chen / Kathrin Hnida / Melissa Ann Graewert / Jan Terje Andersen / Rasmus Iversen / Anne Tuukkanen / Dmitri Svergun / Ludvig M Sollid / ; 要旨: Antibodies to the autoantigen transglutaminase 2 (TG2) are a hallmark of celiac disease. We have studied the interaction between TG2 and an anti-TG2 antibody (679-14-E06) derived from a single gut IgA plasma cell of a celiac disease patient. The antibody recognizes one of four identified epitopes targeted by antibodies of plasma cells of the disease lesion. The binding interface was identified by small angle x-ray scattering, ab initio and rigid body modeling using the known crystal structure of TG2 and the crystal structure of the antibody Fab fragment, which was solved at 2.4 Å resolution. The result was confirmed by testing binding of the antibody to TG2 mutants by ELISA and surface plasmon resonance. TG2 residues Arg-116 and His-134 were identified to be critical for binding of 679-14-E06 as well as other epitope 1 antibodies. In contrast, antibodies directed toward the two other main epitopes (epitopes 2 and 3) were not affected by these mutations. Molecular dynamics simulations suggest interactions of 679-14-E06 with the N-terminal domain of TG2 via the CDR2 and CDR3 loops of the heavy chain and the CDR2 loop of the light chain. In addition there were contacts of the framework 3 region of the heavy chain with the catalytic domain of TG2. The results provide an explanation for the biased usage of certain heavy and light chain gene segments by epitope 1-specific antibodies in celiac disease.

登録者

• Melissa Graewert (EMBL-Hamburg, European Molecular Biology Laboratory (EMBL) - Hamburg outstation, Notkestraße 85, Geb. 25A, 22607 Hamburg, Deutschland, Germany)

モデル

• モデル #310: ; タイプ: atomic / ソフトウェア: CRYSOL / ダミー原子の半径: 1.90 A / カイ2乗値: 1.507984; Omokage検索でこの集合体の類似形状データを探す (詳細)

試料

• 試料: 名称: transglutaminase-2 (TGA2) / 試料濃度: 5.5 mg/ml
• バッファ: 名称: 20 mM Tris 150mM NaCl 1mM EDTA / 濃度: 20.00 mM / pH: 7.2 / 組成: 150mM NaCl, 1mM EDTA

要素 #176

名称: TGA / タイプ: protein / 記述: transglutaminase 2 / 分子量: 79.491 / 分子数: 1 / 由来: Homo sapiens
配列:

MGSSHHHHHH SSGLVPRGSH maeelvlerc dleletngrd hhtadlcrek lvvrrgqpfw ltlhfegrny easvdsltfs vvtgpapsqe agtkarfplr daveegdwta tvvdqqdctl slqlttpana piglyrlsle astgyqgssf vlghfillfn awcpadavyl dseeerqeyv ltqqgfiyqg sakfiknipw nfgqfedgil diclilldvn pkflknagrd csrrsspvyv grvvsgmvnc nddqgvllgr wdnnygdgvs pmswigsvdi lrrwknhgcq rvkygqcwvf aavactvlrc lgiptrvvtn ynsahdqnsn llieyfrnef geiqgdksem iwnfhcwves wmtrpdlqpg yegwqaldpt pqeksegtyc cgpvpvraik egdlstkyda pfvfaevnad vvdwiqqddg svhksinrsl ivglkistks vgrderedit htykypegss eereaftran hlnklaekee tgmamrirvg qsmnmgsdfd vfahitnnta eeyvcrlllc artvsyngil gpecgtkyll nlnlepfsek svplcilyek yrdcltesnl ikvrallvep vinsyllaer dlylenpeik irilgepkqk rklvaevslq nplpvalegc tftvegaglt eeqktveipd pveageevkv rmdllplhmg lhklvvnfes dklkavkgfr nviigpa

実験情報

• ビーム: 設備名称: PETRA III P12 / 地域: Hamburg / 国: Germany / 線源: X-ray synchrotron / 波長: 0.12 Å / スペクトロメータ・検出器間距離: 3.1 mm
• 検出器: 名称: Pilatus 2M

スキャン

タイトル: transglutaminase 2 (TG2 mixture) / 測定日: 2015年1月17日 / 保管温度: 10 °C / セル温度: 10 °C / 照射時間: 0.05 sec. / フレーム数: 20 / 単位: 1/nm /

	Min	Max
Q	0.0199	4.5094

距離分布関数 P(R)

ソフトウェア P(R): GNOM 5.0 / ポイント数: 1096 /

	Min	Max
Q	0.127957	3.0129
P(R) point	1	1096
R	0	12

結果

カーブのタイプ: single_conc
コメント: Structural basis for antigen recognition by transglutaminase 2-specific autoantibodies in celiac disease --- this is the scattering profile of TG2 by itself that is comprised primarilly of monomers and higher oligomeric species (monomer 92%; dimer 8% volume fraction.)

	実験値	Standard	Porod
分子量	77 kDa	77 kDa	73 kDa
体積	-	-	117 nm3

	P(R)	P(R) error	Guinier	Guinier error
前方散乱 I0	1913	15.3	1922	8
慣性半径, Rg	3.43 nm	0.02	3.38 nm	0.02

	Min	Max	Error
D	-	12	0.5
Guinier point	32	139	-