SASDFR8: Bovine serum albumin, purified dimer - SEC-SAXS coupled to multia...

Basic information

• Entry: Database: SASBDB / ID: SASDFR8

Sample

Bovine serum albumin, purified dimer - SEC-SAXS coupled to multiangle laser and quasi-elastic light scattering (MALLS and QELS)

• Bovine serum albumin (protein), BSA, Bos taurus

Function / homology

Function:

enterobactin binding / cellular response to calcium ion starvation / negative regulation of mitochondrial depolarization / toxic substance binding / cellular response to starvation / fatty acid binding / pyridoxal phosphate binding / protein-containing complex / DNA binding / extracellular space / extracellular region / metal ion binding / cytoplasm

Domain/homology:

Serum albumin/Alpha-fetoprotein/Afamin / ALB/AFP/VDB / Serum albumin, N-terminal / Serum albumin, conserved site / Serum albumin-like / Serum albumin family / Albumin domain signature. / Albumin domain profile. / serum albumin

Component:

Albumin

• Biological species: Bos taurus (cattle)

Contact author

• Melissa Graewert
• Cy M Jeffries

Models

• Model #3520: ; Type: dummy / Radius of dummy atoms: 1.90 A / Symmetry: P2 / Chi-square value: 1.05 / P-value: 0.184874; Search similar-shape structures of this assembly by Omokage search (details)
• Model #3524: ; Type: atomic / Radius of dummy atoms: 1.90 A / Chi-square value: 4.375; Search similar-shape structures of this assembly by Omokage search (details)

Sample

• Sample: Name: Bovine serum albumin, purified dimer - SEC-SAXS coupled to multiangle laser and quasi-elastic light scattering (MALLS and QELS); Specimen concentration: 5 mg/ml
• Buffer: Name: 50 mM HEPES, 150 mM NaCl, 2% v/v glycerol, / pH: 7 / Comment: Running buffer for SEC-SAXS

Entity #1309

Name: BSA / Type: protein / Description: Bovine serum albumin / Formula weight: 66.432 / Num. of mol.: 2 / Source: Bos taurus / References: UniProt: P02769
Sequence:

DTHKSEIAHR FKDLGEEHFK GLVLIAFSQY LQQCPFDEHV KLVNELTEFA KTCVADESHA GCEKSLHTLF GDELCKVASL RETYGDMADC CEKQEPERNE CFLSHKDDSP DLPKLKPDPN TLCDEFKADE KKFWGKYLYE IARRHPYFYA PELLYYANKY NGVFQECCQA EDKGACLLPK IETMREKVLA SSARQRLRCA SIQKFGERAL KAWSVARLSQ KFPKAEFVEV TKLVTDLTKV HKECCHGDLL ECADDRADLA KYICDNQDTI SSKLKECCDK PLLEKSHCIA EVEKDAIPEN LPPLTADFAE DKDVCKNYQE AKDAFLGSFL YEYSRRHPEY AVSVLLRLAK EYEATLEECC AKDDPHACYS TVFDKLKHLV DEPQNLIKQN CDQFEKLGEY GFQNALIVRY TRKVPQVSTP TLVEVSRSLG KVGTRCCTKP ESERMPCTED YLSLILNRLC VLHEKTPVSE KVTKCCTESL VNRRPCFSAL TPDETYVPKA FDEKLFTFHA DICTLPDTEK QIKKQTALVE LLKHKPKATE EQLKTVMENF VAFVDKCCAA DDKEACFAVE GPKLVVSTQT ALA

Experimental information

• Beam: Instrument name: PETRA III EMBL P12 / City: Hamburg / 国: Germany / Type of source: X-ray synchrotron / Wavelength: 0.123982 Å / Dist. spec. to detc.: 3 mm
• Detector: Name: Pilatus 6M

Scan

Title: Bovine serum albumin, purified dimer - SEC-SAXS coupled to multiangle laser and quasi-elastic light scattering (MALLS and QELS)
Measurement date: Apr 5, 2019 / Storage temperature: 20 °C / Cell temperature: 20 °C / Exposure time: 1 sec. / Number of frames: 25 / Unit: 1/nm /

	Min	Max
Q	0.1338	6.7984

Distance distribution function P(R)

Sofotware P(R): GNOM 5.0 / Number of points: 1684 /

	Min	Max
Q	0.136558	4.78876
P(R) point	1	1684
R	0	13.2

Result

Type of curve: sec
Comments: BSA dimer underwent pre-purification prior to SEC-SAXS using the following method. All procedures were performed at 4 oC. Protein powder (Sigma) consisting of BSA monomers, dimers, trimers and higher MW species 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 200 Increase 10/300 column (GE Healthcare) equilibrated in the same buffer (flow rate = 0.4 ml/min). Fractionated aliquots corresponding to the minor dimer peak (estimated using UV A280 and UV A245 nm) were pooled and concentrated (30 kDa centrifuge spin filter) to a final concentration of 5 mg/ml (the concentration was determined from triplicate UV A280 measurements using an E0.1% of 0.646 (= 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 4.6 nm.

	Experimental	Porod
MW	126 kDa	132 kDa
Volume	-	211 nm3

	P(R)	Guinier	Guinier error
Forward scattering, I0	12430	12356	9
Radius of gyration, Rg	4.012 nm	3.95 nm	-

	Min	Max
D	-	13.2
Guinier point	1	71