SASDFJ8: Glutamate/aspartate import solute-binding protein (DEBP) in the p...

Basic information

Entry

Database: SASBDB / ID: SASDFJ8

Sample

Glutamate/aspartate import solute-binding protein (DEBP) in the presence of glutamate - Glu-bound 10-fold excess

• Glutamate/aspartate import solute-binding protein (protein), Escherichia coli (strain K12)

Function / homology

Function:

aspartate binding / L-glutamate transmembrane transport / L-aspartate transmembrane transport / L-aspartate import across plasma membrane / L-glutamate import across plasma membrane / glutamate binding / ATP-binding cassette (ABC) transporter complex, substrate-binding subunit-containing / outer membrane-bounded periplasmic space / membrane

Domain/homology:

Bacterial periplasmic substrate-binding proteins / Bacterial extracellular solute-binding proteins, family 3 / Solute-binding protein family 3/N-terminal domain of MltF

Component:

Glutamate/aspartate import solute-binding protein

• Biological species: Escherichia coli (strain K12) (bacteria)
• Citation: Date: 2019 Aug 1; Title: Structure-based screening of binding affinities via small-angle X-ray scattering; Authors: Chen P / Masiewicz P / Perez K

Contact author

• Po-chia Chen (EMBL, European Molecular Biology Laboratory (EMBL) - Heidelberg, Heidelberg, Germany)

Models

Sample

• Sample: Name: Glutamate/aspartate import solute-binding protein (DEBP) in the presence of glutamate - Glu-bound 10-fold excess; Specimen concentration: 2.2 mg/ml
• Buffer: Name: 100 mM NaCl, 20 mM NaPO4, 0.5 mM TCEP / pH: 7.4 / Comment: SEC buffer for PBP experiments

Entity #1755

Type: protein
Description: Glutamate/aspartate import solute-binding protein
Formula weight: 32.052 / Num. of mol.: 1 / Source: Escherichia coli (strain K12) / References: UniProt: P37902
Sequence:

HHHHHHDDAA PAAGSTLDKI AKNGVIVVGH RESSVPFSYY DNQQKVVGYS QDYSNAIVEA VKKKLNKPDL QVKLIPITSQ NRIPLLQNGT FDFECGSTTN NVERQKQAAF SDTIFVVGTR LLTKKGGDIK DFANLKDKAV VVTSGTTSEV LLNKLNEEQK MNMRIISAKD HGDSFRTLES GRAVAFMMDD ALLAGERAKA KKPDNWEIVG KPQSQEAYGC MLRKDDPQFK KLMDDTIAQV QTSGEAEKWF DKWFKNPIPP KNLNMNFELS DEMKALFKEP NDKALN

Experimental information

• Beam: Instrument name: ESRF BM29 / City: Grenoble / 国: France / Type of source: X-ray synchrotronSynchrotron / Wavelength: 0.09919 Å / Dist. spec. to detc.: 2.849 mm
• Detector: Name: Pilatus 1M / Type: Dectris / Pixsize x: 172 mm

Scan

Title: Glutamate/aspartate import solute-binding protein (DEBP) in the presence of glutamate - Glu-bound 10-fold excess
Measurement date: Sep 10, 2018 / Cell temperature: 20 °C / Exposure time: 2 sec. / Number of frames: 12 / Unit: 1/nm /

	Min	Max
Q	0.0306	4.9462

Distance distribution function P(R)

Sofotware P(R): GNOM 5.0 / Number of points: 573 /

	Min	Max
Q	0.101354	2.79976
P(R) point	1	573
R	0	6.44

Result

Type of curve: single_conc
Comments: Glu-bound DEBP at a ligand:protein ratio of 10:1. Ligand condition is prepared by dissolving glutamate in powder form (Sigma-Aldrich) directly into the SAXS buffer, followed by dilution to the target concentration with additional SAXS buffer. Note that the scattering intensity therefore contains Na+ counterions that was not directly subtracted.

	Experimental	Porod
MW	31 kDa	24 kDa
Volume	-	39 nm3

	P(R)	Guinier	Guinier error
Forward scattering, I0	66.97	67.06	0.08
Radius of gyration, Rg	2.061 nm	2.06 nm	0.13

	Min	Max
D	-	6.44
Guinier point	22	128