SASDAX3: MutS tetramer (DNA mismatch repair protein MutS)

Basic information

Entry

Database: SASBDB / ID: SASDAX3

Sample

MutS tetramer

• DNA mismatch repair protein MutS (protein), MutS tetramer, Escherichia coli

Function / homology

Function:

adenine/cytosine mispair binding / MutS complex / mismatch repair complex / regulation of DNA recombination / mismatched DNA binding / DNA binding, bending / ATP-dependent DNA damage sensor activity / mismatch repair / ADP binding / damaged DNA binding / DNA damage response / ATP hydrolysis activity / ATP binding / identical protein binding / cytosol

Domain/homology:

DNA mismatch repair protein MutS / DNA mismatch repair protein MutS/MSH / DNA mismatch repair protein MutS-like, N-terminal / DNA mismatch repair protein MutS, connector domain / DNA mismatch repair protein MutS, clamp / DNA mismatch repair protein MutS, N-terminal / MutS, connector domain superfamily / MutS domain I / MutS domain II / MutS family domain IV / MutS domain III / DNA mismatch repair MutS family / DNA mismatch repair protein MutS, C-terminal / DNA mismatch repair protein MutS, core / DNA mismatch repair protein MutS, core domain superfamily / MutS domain V / DNA mismatch repair proteins mutS family signature. / DNA-binding domain of DNA mismatch repair MUTS family / ATPase domain of DNA mismatch repair MUTS family / P-loop containing nucleoside triphosphate hydrolase

Component:

DNA mismatch repair protein MutS

• Biological species: Escherichia coli (E. coli)
• Citation: Journal: Nucleic Acids Res / Year: 2013; Title: Using stable MutS dimers and tetramers to quantitatively analyze DNA mismatch recognition and sliding clamp formation.; Authors: Flora S Groothuizen / Alexander Fish / Maxim V Petoukhov / Annet Reumer / Laura Manelyte / Herrie H K Winterwerp / Martin G Marinus / Joyce H G Lebbink / Dmitri I Svergun / Peter Friedhoff / Titia K Sixma / ; Abstract: The process of DNA mismatch repair is initiated when MutS recognizes mismatched DNA bases and starts the repair cascade. The Escherichia coli MutS protein exists in an equilibrium between dimers and tetramers, which has compromised biophysical analysis. To uncouple these states, we have generated stable dimers and tetramers, respectively. These proteins allowed kinetic analysis of DNA recognition and structural analysis of the full-length protein by X-ray crystallography and small angle X-ray scattering. Our structural data reveal that the tetramerization domains are flexible with respect to the body of the protein, resulting in mostly extended structures. Tetrameric MutS has a slow dissociation from DNA, which can be due to occasional bending over and binding DNA in its two binding sites. In contrast, the dimer dissociation is faster, primarily dependent on a combination of the type of mismatch and the flanking sequence. In the presence of ATP, we could distinguish two kinetic groups: DNA sequences where MutS forms sliding clamps and those where sliding clamps are not formed efficiently. Interestingly, this inability to undergo a conformational change rather than mismatch affinity is correlated with mismatch repair.

Contact author

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

Models

Sample

• Sample: Name: MutS tetramer / Sample MW: 381 kDa / Specimen concentration: 4.86 mg/ml
• 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 #40

Name: MutS tetramer / Type: protein / Description: DNA mismatch repair protein MutS / Formula weight: 95.25 / Num. of mol.: 4 / Source: Escherichia coli / References: UniProt: P23909
Sequence:

MSAIENFDAH TPMMQQYLRL KAQHPEILLF YRMGDFYELF YDDAKRASQL LDISLTKRGA SAGEPIPMAG IPYHAVENYL AKLVNQGESV AICEQIGDPA TSKGPVERKV VRIVTPGTIS DEALLQERQD NLLAAIWQDS KGFGYATLDI SSGRFRLSEP ADRETMAAEL QRTNPAELLY AEDFAEMSLI EGRRGLRRRP LWEFEIDTAR QQLNLQFGTR DLVGFGVENA PRGLCAAGCL LQYAKDTQRT TLPHIRSITM EREQDSIIMD AATRRNLEIT QNLAGGAENT LASVLDCTVT PMGSRMLKRW LHMPVRDTRV LLERQQTIGA LQDFTAGLQP VLRQVGDLER ILARLALRTA RPRDLARMRH AFQQLPELRA QLETVDSAPV QALREKMGEF AELRDLLERA IIDTPPVLVR DGGVIASGYN EELDEWRALA DGATDYLERL EVRERERTGL DTLKVGFNAV HGYYIQISRG QSHLAPINYM RRQTLKNAER YIIPELKEYE DKVLTSKGKA LALEKQLYEE LFDLLLPHLE ALQQSASALA ELDVLVNLAE RAYTLNYTCP TFIDKPGIRI TEGRHPVVEQ VLNEPFIANP LNLSPQRRML IITGPNMGGK STYMRQTALI ALMAYIGSYV PAQKVEIGPI DRIFTRVGAA DDLASGRSTF MVEMTETANI LHNATEYSLV LMDEIGRGTS TYDGLSLAWA CAENLANKIK ALTLFATHYF ELTQLPEKME GVANVHLDAL EHGDTIAFMH SVQDGAASKS YGLAVAALAG VPKEVIKRAR QKLRELESIS PNAAATQVDG TQMSLLSVPE ETSPAVEALE NLDPDSLTPR QALEWIYRLK SLV

Experimental information

• Beam: Instrument name: DORIS III X33 / City: Hamburg / 国: Germany / Type of source: X-ray synchrotronSynchrotron
• Detector: Name: Pilatus 2M

Scan

Title: MutS, c=4.86 / Measurement date: May 12, 2011 / Storage temperature: 10 °C / Cell temperature: 10 °C / Unit: 1/nm /

	Min	Max
Q	0.0671	6.0241

Distance distribution function P(R)

Sofotware P(R): GNOM 4.5a / Number of points: 397 /

	Min	Max
Q	0.1136	1.197
P(R) point	18	414
R	0	29.18

Result

I0 from Guinier: 301 / Rg from Guinier: 8.5 nm / D max: 29 / Type of curve: single_conc /

	Experimental	Porod
MW	340 kDa	-
Volume	-	750 nm3