SASDDK2: Aspergillus fumigatus UDP galactopyranose mutase

基本情報

• 登録情報: データベース: SASBDB / ID: SASDDK2

試料

Aspergillus fumigatus UDP galactopyranose mutase

• Aspergillus fumigatus UDP galactopyranose mutase (protein)

• 引用: ジャーナル: J Biol Chem / 年: 2012; タイトル: Crystal structures and small-angle x-ray scattering analysis of UDP-galactopyranose mutase from the pathogenic fungus Aspergillus fumigatus.; 著者: Richa Dhatwalia / Harkewal Singh / Michelle Oppenheimer / Dale B Karr / Jay C Nix / Pablo Sobrado / John J Tanner / ; 要旨: UDP-galactopyranose mutase (UGM) is a flavoenzyme that catalyzes the conversion of UDP-galactopyranose to UDP-galactofuranose, which is a central reaction in galactofuranose biosynthesis. Galactofuranose has never been found in humans but is an essential building block of the cell wall and extracellular matrix of many bacteria, fungi, and protozoa. The importance of UGM for the viability of many pathogens and its absence in humans make UGM a potential drug target. Here we report the first crystal structures and small-angle x-ray scattering data for UGM from the fungus Aspergillus fumigatus, the causative agent of aspergillosis. The structures reveal that Aspergillus UGM has several extra secondary and tertiary structural elements that are not found in bacterial UGMs yet are important for substrate recognition and oligomerization. Small-angle x-ray scattering data show that Aspergillus UGM forms a tetramer in solution, which is unprecedented for UGMs. The binding of UDP or the substrate induces profound conformational changes in the enzyme. Two loops on opposite sides of the active site move toward each other by over 10 Å to cover the substrate and create a closed active site. The degree of substrate-induced conformational change exceeds that of bacterial UGMs and is a direct consequence of the unique quaternary structure of Aspergillus UGM. Galactopyranose binds at the re face of the FAD isoalloxazine with the anomeric carbon atom poised for nucleophilic attack by the FAD N5 atom. The structural data provide new insight into substrate recognition and the catalytic mechanism and thus will aid inhibitor design.

登録者

• John Tanner (Mizzou, University of Missouri-Columbia, Columbia, MO, USA)

モデル

• モデル #1716: ; タイプ: atomic / ダミー原子の半径: 1.90 A / カイ2乗値: 12.0390640394; 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: Aspergillus fumigatus UDP galactopyranose mutase / 試料濃度: 9 mg/ml
• バッファ: 名称: 20 mM HEPES, 45 mM NaCl, 0.5 mM Tris(hydroxypropyl)phosphine; pH: 7.5

要素 #930

タイプ: protein / 記述: Aspergillus fumigatus UDP galactopyranose mutase / 分子量: 57.068 / 分子数: 4
配列:

AIAMTHPDIS VDVLVIGAGP TGLGAAKRLN QIDGPSWMIV DSNETPGGLA STDVTPEGFL YDVGGHVIFS HYKYFDDCLD EALPKEDDWY THQRISYVRC QGQWVPYPFQ NNISMLPKEE QVKCIDGMID AALEARVANT KPKTFDEWIV RMMGTGIADL FMRPYNFKVW AVPTTKMQCA WLGERVAAPN LKAVTTNVIL GKTAGNWGPN ATFRFPARGG TGGIWIAVAN TLPKEKTRFG EKGKVTKVNA NNKTVTLQDG TTIGYKKLVS TMAVDFLAEA MNDQELVGLT KQLFYSSTHV IGVGVRGSRP ERIGDKCWLY FPEDNCPFYR ATIFSNYSPY NQPEASAALP TMQLADGSRP QSTEAKEGPY WSIMLEVSES SMKPVNQETI LADCIQGLVN TEMLKPTDEI VSTYHRRFDH GYPTPTLERE GTLTQILPKL QDKDIWSRGR FGSWRYEVGN QDHSFMLGVE AVDNIVNGAV ELTLNYPDFV NGRQNTERRL VDGAQVFAKS KAQ

実験情報

• ビーム: 設備名称: Advanced Light Source (ALS) 12.3.1 (SIBYLS) / 地域: Berkeley, CA / 国: USA / 線源: X-ray synchrotron / 波長: 0.1127 Å
• 検出器: 名称: MAR 165 CCD

スキャン

タイトル: Aspergillus fumigatus UDP galactopyranose mutase
測定日: 2010年4月19日 / セル温度: 10 °C / 単位: 1/A /

	Min	Max
Q	0.0106	0.3213

距離分布関数 P(R)

ソフトウェア P(R): GNOM 5.0 / ポイント数: 512 /

	Min	Max
Q	0.010641	0.321334
P(R) point	1	512
R	0	146.8

結果

カーブのタイプ: single_conc /

	実験値	Porod
分子量	228.3 kDa	-
体積	-	308 nm3

	P(R)	P(R) error	Guinier	Guinier error
前方散乱 I0	893.5	1	892.91	1.7
慣性半径, Rg	4.77 nm	-	4.727 nm	0.013

	Min	Max
D	-	14.68
Guinier point	1	28