2gb7

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{{STRUCTURE_2gb7| PDB=2gb7 | SCENE= }}
{{STRUCTURE_2gb7| PDB=2gb7 | SCENE= }}
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'''Metal-depleted Ecl18kI in complex with uncleaved, modified DNA'''
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===Metal-depleted Ecl18kI in complex with uncleaved, modified DNA===
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==Overview==
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Restricion endonuclease Ecl18kI is specific for the sequence /CCNGG and cleaves it before the outer C to generate 5 nt 5'-overhangs. It has been suggested that Ecl18kI is evolutionarily related to NgoMIV, a 6-bp cutter that cleaves the sequence G/CCGGC and leaves 4 nt 5'-overhangs. Here, we report the crystal structure of the Ecl18kI-DNA complex at 1.7 A resolution and compare it with the known structure of the NgoMIV-DNA complex. We find that Ecl18kI flips both central nucleotides within the CCNGG sequence and buries the extruded bases in pockets within the protein. Nucleotide flipping disrupts Watson-Crick base pairing, induces a kink in the DNA and shifts the DNA register by 1 bp, making the distances between scissile phosphates in the Ecl18kI and NgoMIV cocrystal structures nearly identical. Therefore, the two enzymes can use a conserved DNA recognition module, yet recognize different sequences, and form superimposable dimers, yet generate different cleavage patterns. Hence, Ecl18kI is the first example of a restriction endonuclease that flips nucleotides to achieve specificity for its recognition site.
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{{ABSTRACT_PUBMED_16628220}}
==About this Structure==
==About this Structure==
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==Reference==
==Reference==
Nucleotide flips determine the specificity of the Ecl18kI restriction endonuclease., Bochtler M, Szczepanowski RH, Tamulaitis G, Grazulis S, Czapinska H, Manakova E, Siksnys V, EMBO J. 2006 May 17;25(10):2219-29. Epub 2006 Apr 20. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/16628220 16628220]
Nucleotide flips determine the specificity of the Ecl18kI restriction endonuclease., Bochtler M, Szczepanowski RH, Tamulaitis G, Grazulis S, Czapinska H, Manakova E, Siksnys V, EMBO J. 2006 May 17;25(10):2219-29. Epub 2006 Apr 20. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/16628220 16628220]
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Alternative arrangements of catalytic residues at the active sites of restriction enzymes., Tamulaitis G, Solonin AS, Siksnys V, FEBS Lett. 2002 May 8;518(1-3):17-22. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/11997010 11997010]
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The Ecl18kI restriction-modification system: cloning, expression, properties of the purified enzymes., Denjmukhametov MM, Brevnov MG, Zakharova MV, Repyk AV, Solonin AS, Petrauskene OV, Gromova ES, FEBS Lett. 1998 Aug 21;433(3):233-6. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/9744801 9744801]
[[Category: Enterobacter cloacae]]
[[Category: Enterobacter cloacae]]
[[Category: Single protein]]
[[Category: Single protein]]
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[[Category: Nucleotide flipping]]
[[Category: Nucleotide flipping]]
[[Category: Type ii restriction endonuclease]]
[[Category: Type ii restriction endonuclease]]
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Tue Jul 29 15:33:34 2008''

Revision as of 12:33, 29 July 2008

Image:2gb7.png

Template:STRUCTURE 2gb7

Metal-depleted Ecl18kI in complex with uncleaved, modified DNA

Template:ABSTRACT PUBMED 16628220

About this Structure

2GB7 is a Single protein structure of sequence from Enterobacter cloacae. Full crystallographic information is available from OCA.

Reference

Nucleotide flips determine the specificity of the Ecl18kI restriction endonuclease., Bochtler M, Szczepanowski RH, Tamulaitis G, Grazulis S, Czapinska H, Manakova E, Siksnys V, EMBO J. 2006 May 17;25(10):2219-29. Epub 2006 Apr 20. PMID:16628220

Alternative arrangements of catalytic residues at the active sites of restriction enzymes., Tamulaitis G, Solonin AS, Siksnys V, FEBS Lett. 2002 May 8;518(1-3):17-22. PMID:11997010

The Ecl18kI restriction-modification system: cloning, expression, properties of the purified enzymes., Denjmukhametov MM, Brevnov MG, Zakharova MV, Repyk AV, Solonin AS, Petrauskene OV, Gromova ES, FEBS Lett. 1998 Aug 21;433(3):233-6. PMID:9744801

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