SASDCF3: Nucleolysin TIA-1 isoform p40 (TIA-1 wild type)

Basic information

Entry

Database: SASBDB / ID: SASDCF3

Sample

Nucleolysin TIA-1 isoform p40

• Nucleolysin TIA-1 isoform p40 (protein), TIA-1 wild type, Homo sapiens

• Function / homology: Isoform Short of Cytotoxic granule associated RNA binding protein TIA1
• Biological species: Homo sapiens (human)
• Citation: Journal: Angew Chem Int Ed Engl / Year: 2017; Title: Segmental, Domain-Selective Perdeuteration and Small-Angle Neutron Scattering for Structural Analysis of Multi-Domain Proteins.; Authors: Miriam Sonntag / Pravin Kumar Ankush Jagtap / Bernd Simon / Marie-Sousai Appavou / Arie Geerlof / Ralf Stehle / Frank Gabel / Janosch Hennig / Michael Sattler / ; Abstract: Multi-domain proteins play critical roles in fine-tuning essential processes in cellular signaling and gene regulation. Typically, multiple globular domains that are connected by flexible linkers undergo dynamic rearrangements upon binding to protein, DNA or RNA ligands. RNA binding proteins (RBPs) represent an important class of multi-domain proteins, which regulate gene expression by recognizing linear or structured RNA sequence motifs. Here, we employ segmental perdeuteration of the three RNA recognition motif (RRM) domains in the RBP TIA-1 using Sortase A mediated protein ligation. We show that domain-selective perdeuteration combined with contrast-matched small-angle neutron scattering (SANS), SAXS and computational modeling provides valuable information to precisely define relative domain arrangements. The approach is generally applicable to study conformational arrangements of individual domains in multi-domain proteins and changes induced by ligand binding.

Contact author

• Miriam Sonntag (Technical University Munich)

Models

Sample

• Sample: Name: Nucleolysin TIA-1 isoform p40 / Specimen concentration: 2.50-5.00
• Buffer: Name: 10 mM Potassium Phosphate 50 mM NaCl 10 mM DTT / pH: 6

Entity #659

Name: TIA-1 wild type / Type: protein / Description: Nucleolysin TIA-1 isoform p40 / Formula weight: 30.457 / Num. of mol.: 1 / Source: Homo sapiens / References: UniProt: P31483-2
Sequence:

MEDEMPKTLY VGNLSRDVTE ALILQLFSQI GPCKNCKMIM DTAGNDPYCF VEFHEHRHAA AALAAMNGRK IMGKEVKVNW ATTPSSQKKD TSNHFHVFVG DLSPEITTED IKAAFAPFGR ISDARVVKDM ATGKSKGYGF VSFFNKWDAE NAIQQMGGQW LGGRQIRTNW ATRKPPAPKS TYESNTKQLS YDEVVNQSSP SNCTVYCGGV TSGLTEQLMR QTFSPFGQIM EIRVFPDKGY SFVRFNSHES AAHAIVSVNG TTIEGHVVKC YWGK

Experimental information

• Beam: Instrument name: Rigaku BioSAXS-1000 / City: Munich / 国: Germany / Type of source: X-ray in house / Wavelength: 0.1542 Å / Dist. spec. to detc.: 0.482 mm
• Detector: Name: Pilatus 100K / Pixsize x: 172 mm

Scan

Title: Nucleolysin TIA-1 isoform p40 / Measurement date: Apr 28, 2016 / Storage temperature: 20 °C / Cell temperature: 20 °C / Exposure time: 7200 sec. / Number of frames: 8 / Unit: 1/A /

	Min	Max
Q	0.0209	0.7056

Distance distribution function P(R)

Sofotware P(R): GNOM 5.0 / Number of points: 159 /

	Min	Max
Q	0.026587	0.252454
P(R) point	1	159
R	0	111.7

Result

Type of curve: merged

	Experimental	Standard	Porod
MW	29.9 kDa	29.9 kDa	29.8 kDa
Volume	-	-	38 nm3

	P(R)	P(R) error	Guinier	Guinier error
Forward scattering, I0	0.0237	0.0002	0.023	0.0002
Radius of gyration, Rg	2.92 nm	0.04	2.73 nm	0.03

	Min	Max
D	-	11.2
Guinier point	5	22