SASDCN9: Ks-Amt5-HK (apo) (Ammonium transporter histidine kinase domain, K...

Basic information

Entry

Database: SASBDB / ID: SASDCN9

Sample

Ks-Amt5-HK (apo)

• Ammonium transporter histidine kinase domain (protein), Ks-Amt5-HK, Candidatus Kuenenia stuttgartiensis

Function / homology

Function:

ammonium homeostasis / ammonium channel activity / phosphorelay sensor kinase activity / plasma membrane

Domain/homology:

Ammonium transporter / Ammonium transporter AmtB-like domain / Ammonium Transporter Family / Ammonium/urea transporter / His Kinase A (phospho-acceptor) domain / His Kinase A (phosphoacceptor) domain / Signal transduction histidine kinase, dimerisation/phosphoacceptor domain / Signal transduction histidine kinase, dimerisation/phosphoacceptor domain superfamily / Signal transduction histidine kinase-related protein, C-terminal / Histidine kinase domain / Histidine kinase domain profile. / Histidine kinase-, DNA gyrase B-, and HSP90-like ATPase / Histidine kinase-like ATPases / Histidine kinase/HSP90-like ATPase / Histidine kinase/HSP90-like ATPase superfamily

Component:

Ammonium transporter

• Biological species: Candidatus Kuenenia stuttgartiensis (bacteria)
• Citation: Journal: Nat Commun / Year: 2018; Title: Signaling ammonium across membranes through an ammonium sensor histidine kinase.; Authors: Tobias Pflüger / Camila F Hernández / Philipp Lewe / Fabian Frank / Haydyn Mertens / Dmitri Svergun / Manfred W Baumstark / Vladimir Y Lunin / Mike S M Jetten / Susana L A Andrade / ; Abstract: Sensing and uptake of external ammonium is essential for anaerobic ammonium-oxidizing (anammox) bacteria, and is typically the domain of the ubiquitous Amt/Rh ammonium transporters. Here, we report on the structure and function of an ammonium sensor/transducer from the anammox bacterium "Candidatus Kuenenia stuttgartiensis" that combines a membrane-integral ammonium transporter domain with a fused histidine kinase. It contains a high-affinity ammonium binding site not present in assimilatory Amt proteins. The levels of phosphorylated histidine in the kinase are coupled to the presence of ammonium, as conformational changes during signal recognition by the Amt module are transduced internally to modulate the kinase activity. The structural analysis of this ammonium sensor by X-ray crystallography and small-angle X-ray-scattering reveals a flexible, bipartite system that recruits a large uptake transporter as a sensory module and modulates its functionality to achieve a mechanistic coupling to a kinase domain in order to trigger downstream signaling events.

Contact author

• Haydyn Mertens (EMBL-Hamburg, European Molecular Biology Laboratory (EMBL) - Hamburg outstation, Notkestraße 85, Geb. 25A, 22607 Hamburg, Deutschland, Germany)

Models

Sample

• Sample: Name: Ks-Amt5-HK (apo) / Specimen concentration: 1.00-5.00
• Buffer: Name: 20 mM Tris/HCl,100 mM NaCl, 5%(w/v) glycerol / pH: 8

Entity #910

Name: Ks-Amt5-HK / Type: protein / Description: Ammonium transporter histidine kinase domain / Formula weight: 30.031 / Num. of mol.: 1 / Source: Candidatus Kuenenia stuttgartiensis / References: UniProt: Q1Q357
Sequence:

FIRITRLQDI NTILEKRVQE KTADLQMANV ALEKANRLKS EFLTTMSAEL RTPLNAIIGF AEVLRDEIAG SLSKDQKEYV TDIHSSGHHL LDMINNILDL SKIETGKMHL QYEEFCIEDA INDTLTIINA SANNKGISVH TNIQDNTPLL SADKTKFRQI LYNLLSNAVK FTPENGKITI NVFQKDNSLQ FEIVDTGIGI KPEDKEKLFE AFHQADASLT REYEGTGLGL HLTKRLVELH GGKIWAESTF GKGSTFFFIL PINPVNK

Experimental information

• Beam: Instrument name: PETRA III P12 / City: Hamburg / 国: Germany / Type of source: X-ray synchrotronSynchrotron / Wavelength: 0.124 Å / Dist. spec. to detc.: 2.7 mm
• Detector: Name: Pilatus 2M

Scan

Title: Ks-Amt5-HK (apo) / Measurement date: Mar 2, 2013 / Storage temperature: 20 °C / Cell temperature: 20 °C / Exposure time: 0.05 sec. / Number of frames: 20 / Unit: 1/nm /

	Min	Max
Q	0.145	4.423

Distance distribution function P(R)

Sofotware P(R): GNOM 5.0 / Number of points: 1200 /

	Min	Max
Q	0.187424	3.37004
P(R) point	1	1200
R	0	9

Result

Type of curve: merged

	Experimental	Standard	Standard error	Porod
MW	32.6 kDa	32.6 kDa	3	42 kDa
Volume	-	-	-	71 nm3

	P(R)	P(R) error	Guinier	Guinier error
Forward scattering, I0	2065	7	2046	9
Radius of gyration, Rg	2.8 nm	1	2.7 nm	0.1

	Min	Max	Error
D	-	10	0.5
Guinier point	2	127	-