SASDFG8: Glutamine-binding periplasmic protein with hexahistidine tag (Gln...

Basic information

Entry

Database: SASBDB / ID: SASDFG8

Sample

Glutamine-binding periplasmic protein with hexahistidine tag (GlnBP) in the presence of glutamine - Gln-bound 10-fold excess

• Glutamine-binding periplasmic protein with hexahistidine tag (protein), GlnBP, Escherichia coli (strain K12)

Function / homology

Function:

glutamine binding / L-glutamine import across plasma membrane / ligand-gated monoatomic ion channel activity / ATP-binding cassette (ABC) transporter complex, substrate-binding subunit-containing / outer membrane-bounded periplasmic space / membrane

Domain/homology:

Glutamine-binding periplasmic protein GlnH, type 2 periplasmic binding protein fold / Solute-binding protein family 3, conserved site / Bacterial extracellular solute-binding proteins, family 3 signature. / Bacterial periplasmic substrate-binding proteins / Bacterial extracellular solute-binding proteins, family 3 / Solute-binding protein family 3/N-terminal domain of MltF / Ionotropic glutamate receptor / Eukaryotic homologues of bacterial periplasmic substrate binding proteins.

Component:

Glutamine-binding periplasmic 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: Glutamine-binding periplasmic protein with hexahistidine tag (GlnBP) in the presence of glutamine - Gln-bound 10-fold excess; Specimen concentration: 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 #1754

Name: GlnBP / Type: protein
Description: Glutamine-binding periplasmic protein with hexahistidine tag
Formula weight: 25.786 / Num. of mol.: 1 / Source: Escherichia coli (strain K12) / References: UniProt: P0AEQ3
Sequence:

HHHHHHADKK LVVATDTAFV PFEFKQGDKY VGFDVDLWAA IAKELKLDYE LKPMDFSGII PALQTKNVDL ALAGITITDE RKKAIDFSDG YYKSGLLVMV KANNNDVKSV KDLDGKVVAV KSGTGSVDYA KANIKTKDLR QFPNIDNAYM ELGTNRADAV LHDTPNILYF IKTAGNGQFK AVGDSLEAQQ YGIAFPKGSD ELRDKVNGAL KTLRENGTYN EIYKKWFGTE PK

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: Glutamine-binding periplasmic protein with hexahistidine tag (GlnBP) in the presence of glutamine - Gln-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	6.1

Result

Type of curve: single_conc
Comments: Gln-bound GlnBP at a ligand:protein ratio of 10:1. Ligand condition is prepared by dissolving glutamine in powder form (Sigma-Aldrich) directly into the SAXS buffer, followed by dilution to the target concentration with additional SAXS buffer.

	Experimental	Porod
MW	25 kDa	21 kDa
Volume	-	34 nm3

	P(R)	Guinier	Guinier error
Forward scattering, I0	49.47	49.65	0.06
Radius of gyration, Rg	2.023 nm	2.03 nm	0.13

	Min	Max
D	-	6.1
Guinier point	15	130