SASDAJ5: Annexin-A4

Basic information

• Entry: Database: SASBDB / ID: SASDAJ5

Sample

Annexin-A4

• Annexin-A4 (protein), Annexin-A4, Homo sapiens

• Biological species: Homo sapiens (human)
• Citation: Journal: Biophys J / Year: 2012; Title: Conformational analysis of a genetically encoded FRET biosensor by SAXS.; Authors: Haydyn D T Mertens / Alen Piljić / Carsten Schultz / Dmitri I Svergun / ; Abstract: Genetically encoded FRET (Foerster resonance energy transfer) sensors are exciting tools in modern cell biology. Changes in the conformation of a sensor lead to an altered emission ratio and provide the means to determine both temporal and spatial changes in target molecules, as well as the activity of enzymes. FRET sensors are widely used to follow phosphorylation events and to monitor the effects of elevated calcium levels. Here, we report for the first time, to our knowledge, on the analysis of the conformational changes involved in sensor function at low resolution using a combination of in vitro and in cellulo FRET measurements and small-angle scattering of x rays (SAXS). The large and dynamic structural rearrangements involved in the modification of the calcium- and phosphorylation-sensitive probe CYNEX4 are comprehensively characterized. It is demonstrated that the synergistic use of SAXS and FRET methods allows one to resolve the ambiguities arising due to the rotation of the sensor molecules and the flexibility of the probe.

Contact author

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

Models

• Model #118: ; Type: atomic / Software: crysol / Chi-square value: 4.289041; Search similar-shape structures of this assembly by Omokage search (details)
• Model #120: ; Type: dummy / Software: dammif / Radius of dummy atoms: 1.90 A / Chi-square value: 5.221225; Search similar-shape structures of this assembly by Omokage search (details)

Sample

• Sample: Name: Annexin-A4 / Sample MW: 36 kDa / Specimen concentration: 0.40-7.30 / Concentration method: A280
• Buffer: Name: HEPES / Concentration: 50.00 mM / PK: 7 / pH: 7.5 / Comment: 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid / Composition: KCl 50.000 mM

Entity #92

Name: Annexin-A4 / Type: protein / Description: Annexin-A4 / Formula weight: 36 / Num. of mol.: 1 / Source: Homo sapiens
Sequence:

MAMATKGGTV KAASGFNAME DAQTLRKAMK GLGTDEDAII SVLAYRNTAQ RQEIRTAYKS TIGRDLIDDL KSELSGNFEQ VIVGMMTPTV LYDVQELRRA MKGAGTDEGC LIEILASRTP EEIRRISQTY QQQYGRSLED DIRSDTSFMF QRVLVSLSAG GRDEGNYLDD ALVRQDAQDL YEAGEKKWGT DEVKFLTVLC SRNRNHLLHV FDEYKRISQK DIEQSIKSET SGSFEDALLA IVKCMRNKSA YFAEKLYKSM KGLGTDDNTL IRVMVSRAEI DMLDIRAHFK RLYGKSLYSF IKGDTSGDYR KVLLVLCGGD D

Experimental information

• Beam: Instrument name: DORIS III X33 / City: Hamburg / 国: Germany / Shape: 0.6 / Type of source: X-ray synchrotron / Wavelength: 0.15 Å / Dist. spec. to detc.: 2.7 mm
• Detector: Name: Pilatus 1M-W / Pixsize x: 0.172 mm

Scan

Title: Annexin-A4 / Measurement date: Sep 11, 2010 / Storage temperature: 10 °C / Cell temperature: 10 °C / Exposure time: 30 sec. / Number of frames: 4 / Unit: 1/nm /

	Min	Max
Q	0.0731	5.713

Distance distribution function P(R)

Sofotware P(R): GNOM 4.6 / Number of points: 399 /

	Min	Max
Q	0.2436	3.33
P(R) point	66	464
R	0	7.6

Result

Type of curve: extrapolated / Standard: BSA /

	Experimental	Standard	Porod
MW	33 kDa	33 kDa	-
Volume	-	-	54 nm3

	P(R)	Guinier
Forward scattering, I0	39.9	40.26
Radius of gyration, Rg	2.4 nm	2.4 nm

	Min	Max
D	-	7.6
Guinier point	66	183