SASDFP8: Carbonic anhydrase 2 from bovine erythrocytes - SEC-SAXS coupled ...

基本情報

• 登録情報: データベース: SASBDB / ID: SASDFP8

試料

Carbonic anhydrase 2 from bovine erythrocytes - SEC-SAXS coupled to multiangle laser and quasi-elastic light scattering (MALLS and QELS)

• Carbonic anhydrase 2 (protein), CA2, Bos taurus

機能・相同性

分子機能:

positive regulation of dipeptide transmembrane transport / regulation of monoatomic anion transport / cyanamide hydratase / cyanamide hydratase activity / angiotensin-activated signaling pathway / regulation of intracellular pH / carbonic anhydrase / carbonate dehydratase activity / carbon dioxide transport / apical part of cell / zinc ion binding / plasma membrane / cytoplasm

ドメイン・相同性:

Carbonic anhydrase, alpha-class, conserved site / Alpha-carbonic anhydrases signature. / Carbonic anhydrase, alpha-class / Alpha carbonic anhydrase domain / Alpha carbonic anhydrase domain superfamily / Eukaryotic-type carbonic anhydrase / Alpha-carbonic anhydrases profile. / Eukaryotic-type carbonic anhydrase

構成要素:

Carbonic anhydrase 2

• 生物種: Bos taurus (ウシ)

登録者

• Melissa Graewert
• Cy M Jeffries

モデル

• モデル #3213: ; タイプ: dummy / ソフトウェア: (SUPCOMB 23 (r9988)) / ダミー原子の半径: 1.50 A / 対称性: P1; コメント: Refined DAMMIN model obtained from the spatial alignment of 10 individual models (DAMSTART); カイ2乗値: 1.020 / P-value: 0.448891; Omokage検索でこの集合体の類似形状データを探す (詳細)
• モデル #3214: ; タイプ: dummy / ソフトウェア: (DAMFILT 5.0 (r10552)) / ダミー原子の半径: 1.50 A / 対称性: P1; コメント: DAMFILT spatially aligned and volume occupancy corrected (averaged) model from 10 individual models; カイ2乗値: 1.020 / P-value: 0.448891; Omokage検索でこの集合体の類似形状データを探す (詳細)
• モデル #3215: ; タイプ: atomic / 対称性: P1; コメント: Model-fit calculated using 30 harmonics and 300 points; カイ2乗値: 1.095 / P-value: 0.269894; 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: Carbonic anhydrase 2 from bovine erythrocytes - SEC-SAXS coupled to multiangle laser and quasi-elastic light scattering (MALLS and QELS); 試料濃度: 11.9 mg/ml
• バッファ: 名称: 50 mM HEPES, 150 mM NaCl, 2% v/v glycerol, / pH: 7 / コメント: Running buffer for SEC-SAXS

要素 #1768

名称: CA2 / タイプ: protein / 記述: Carbonic anhydrase 2 / 分子量: 29.113 / 分子数: 1 / 由来: Bos taurus / 参照: UniProt: P00921
配列:

MSHHWGYGKH NGPEHWHKDF PIANGERQSP VDIDTKAVVQ DPALKPLALV YGEATSRRMV NNGHSFNVEY DDSQDKAVLK DGPLTGTYRL VQFHFHWGSS DDQGSEHTVD RKKYAAELHL VHWNTKYGDF GTAAQQPDGL AVVGVFLKVG DANPALQKVL DALDSIKTKG KSTDFPNFDP GSLLPNVLDY WTYPGSLTTP PLLESVTWIV LKEPISVSSQ QMLKFRTLNF NAEGEPELLM LANWRPAQPL KNRQVRGFPK

実験情報

• ビーム: 設備名称: PETRA III EMBL P12 / 地域: Hamburg / 国: Germany / 線源: X-ray synchrotron / 波長: 0.123982 Å / スペクトロメータ・検出器間距離: 3 mm
• 検出器: 名称: Pilatus 6M

スキャン

タイトル: Carbonic anhydrase 2 from bovine erythrocytes - SEC-SAXS coupled to multiangle laser and quasi-elastic light scattering (MALLS and QELS)
測定日: 2019年4月5日 / 保管温度: 20 °C / セル温度: 20 °C / 照射時間: 1 sec. / フレーム数: 41 / 単位: 1/nm /

	Min	Max
Q	0.073	7.213

距離分布関数 P(R)

ソフトウェア P(R): GNOM 5.0 / ポイント数: 1784 /

	Min	Max
Q	0.072981	5.00161
P(R) point	1	1784
R	0	5.05

結果

カーブのタイプ: sec
コメント: Carbonic anhydrase underwent pre-purification prior to SEC-SAXS using the following method. All procedures were performed at 4 oC. The protein (from Sigma; Gel Filtration Markers Kit MWGF1000) was made to approximately 25 mg/ml in 25 mM HEPES, 50 mM NaCl, 5 mM urea, 1% v/v glycerol, pH 7. Approximately 200 μl of sample were loaded onto a Superdex 75 Increase 10/300 column (GE Healthcare) equilibrated in the same buffer (flow rate = 0.4 ml/min). Fractionated aliquots corresponding to the highest absorbing peak (estimated using UV A280 and UV A245 nm) were pooled and concentrated (3 kDa centrifuge spin filter) to a final concentration of 11.9 mg/ml (the concentration was determined from triplicate UV A280 measurements using an E0.1% of 1.732 (= 1 g/l) calculated from the amino acid sequence (ProtParam)). Approximately 50 μl aliquots were snap-frozen in liquid nitrogen then stored at -80oC prior to the SEC-SAXS analysis that was performed at room temperature in 50 mM HEPES, 150 mM NaCl, 2% v/v glycerol, pH 7. The Rg-correlation through the SEC-SAXS peak, the individual unsubtracted SEC-SAXS frames as well as the results from coupled MALLS and QELS analysis are included in the full entry zip archive. The quoted experimental molecular weight was determined using MALLS in combination with refractive-index (RI) measurements that were recorded from the same sample eluting from the column using a split-flow SEC-SAXS-light scattering configuration (Graewert et al., (2015) Sci. Reports. 5, 10734: doi: 10.1038/srep10734). The average hydrodynamic radius of the protein is 2.4 nm.

	実験値	Porod
分子量	28.6 kDa	23 kDa
体積	-	36.7 nm3

	P(R)	P(R) error	Guinier	Guinier error
前方散乱 I0	7744	2.5	7753.8	3.15
慣性半径, Rg	1.792 nm	0.001	1.82 nm	-

	Min	Max
D	-	5.05
Guinier point	1	231