Purification and properties of a DNA ligase from a soluble DNA replication complex
A DNA ligase has been purified from a subnuclear soluble replication complex isolated from adenovirus type 2-infected human KB cells. DNA ligase activity could not be demonstrated using an exogenous template until the complex was dissociated, suggesting that the ligase activity may be a component of...
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| Format: |
E-Artikel |
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| Sprache: |
Englisch |
| Erschienen: |
1980 |
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| Reproduktion: |
Elsevier Journal Backfiles on ScienceDirect 1907 - 2002 |
|---|---|
| Übergeordnetes Werk: |
in: BBA Section Nucleic Acids And Protein Synthesis - Amsterdam : Elsevier, 606(1980), 2, Seite 202-213 |
| Übergeordnetes Werk: |
volume:606 ; year:1980 ; number:2 ; pages:202-213 |
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| 520 | |a A DNA ligase has been purified from a subnuclear soluble replication complex isolated from adenovirus type 2-infected human KB cells. DNA ligase activity could not be demonstrated using an exogenous template until the complex was dissociated, suggesting that the ligase activity may be a component of the complex. The purified enzyme was free of endonuclease, exonuclease, 5'-nucleotidase, and phosphatase activites, and had a molecular weight of 105 000, as estimated by sedimentation in a glycerol gradient. The ligase requires ATP and a divalent cation for activity. The optimum of the reaction is at pH 7.8 in 50-100 mM Tris-HCl buffer and 10-20 mM MgCl"2. Monovalent salts greatly stimulate ligase activity and the optimum was found at 150 mM. The reaction is very sensitive to high temperature; maximum activity was observed at 25-30^oC. ATP is the sole required cofactor and NAD, dATP and GTP could not replace the requirement for ATP. The K"m for ATP is 60 μM. The K"m for DNA is 250 μg/ml or 1.6 nmol of terminal phosphate/ml and thus the enzyme shows relatively weak affinity for exogenous DNA. The maximum conversion of ^3^2P into a phosphatase-resistant form is approximately 1.3% of the total, whereas T4 ligase, under the same conditions, can convert more than 25% of phosphate into a resistant form. | ||
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(DE-627)NLEJ185895921 (DE-599)GBVNLZ185895921 DE-627 ger DE-627 rakwb eng Purification and properties of a DNA ligase from a soluble DNA replication complex 1980 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier A DNA ligase has been purified from a subnuclear soluble replication complex isolated from adenovirus type 2-infected human KB cells. DNA ligase activity could not be demonstrated using an exogenous template until the complex was dissociated, suggesting that the ligase activity may be a component of the complex. The purified enzyme was free of endonuclease, exonuclease, 5'-nucleotidase, and phosphatase activites, and had a molecular weight of 105 000, as estimated by sedimentation in a glycerol gradient. The ligase requires ATP and a divalent cation for activity. The optimum of the reaction is at pH 7.8 in 50-100 mM Tris-HCl buffer and 10-20 mM MgCl"2. Monovalent salts greatly stimulate ligase activity and the optimum was found at 150 mM. The reaction is very sensitive to high temperature; maximum activity was observed at 25-30^oC. ATP is the sole required cofactor and NAD, dATP and GTP could not replace the requirement for ATP. The K"m for ATP is 60 μM. The K"m for DNA is 250 μg/ml or 1.6 nmol of terminal phosphate/ml and thus the enzyme shows relatively weak affinity for exogenous DNA. The maximum conversion of ^3^2P into a phosphatase-resistant form is approximately 1.3% of the total, whereas T4 ligase, under the same conditions, can convert more than 25% of phosphate into a resistant form. Elsevier Journal Backfiles on ScienceDirect 1907 - 2002 Tsuruo, T. oth Arens, M. oth Padmanabhan, R. oth Green, M. oth in BBA Section Nucleic Acids And Protein Synthesis Amsterdam : Elsevier 606(1980), 2, Seite 202-213 (DE-627)NLEJ185870457 (DE-600)2209441-6 0005-2787 nnns volume:606 year:1980 number:2 pages:202-213 http://linkinghub.elsevier.com/retrieve/pii/0005-2787(80)90030-1 GBV_USEFLAG_H ZDB-1-SDJ GBV_NL_ARTICLE AR 606 1980 2 202-213 |
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(DE-627)NLEJ185895921 (DE-599)GBVNLZ185895921 DE-627 ger DE-627 rakwb eng Purification and properties of a DNA ligase from a soluble DNA replication complex 1980 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier A DNA ligase has been purified from a subnuclear soluble replication complex isolated from adenovirus type 2-infected human KB cells. DNA ligase activity could not be demonstrated using an exogenous template until the complex was dissociated, suggesting that the ligase activity may be a component of the complex. The purified enzyme was free of endonuclease, exonuclease, 5'-nucleotidase, and phosphatase activites, and had a molecular weight of 105 000, as estimated by sedimentation in a glycerol gradient. The ligase requires ATP and a divalent cation for activity. The optimum of the reaction is at pH 7.8 in 50-100 mM Tris-HCl buffer and 10-20 mM MgCl"2. Monovalent salts greatly stimulate ligase activity and the optimum was found at 150 mM. The reaction is very sensitive to high temperature; maximum activity was observed at 25-30^oC. ATP is the sole required cofactor and NAD, dATP and GTP could not replace the requirement for ATP. The K"m for ATP is 60 μM. The K"m for DNA is 250 μg/ml or 1.6 nmol of terminal phosphate/ml and thus the enzyme shows relatively weak affinity for exogenous DNA. The maximum conversion of ^3^2P into a phosphatase-resistant form is approximately 1.3% of the total, whereas T4 ligase, under the same conditions, can convert more than 25% of phosphate into a resistant form. Elsevier Journal Backfiles on ScienceDirect 1907 - 2002 Tsuruo, T. oth Arens, M. oth Padmanabhan, R. oth Green, M. oth in BBA Section Nucleic Acids And Protein Synthesis Amsterdam : Elsevier 606(1980), 2, Seite 202-213 (DE-627)NLEJ185870457 (DE-600)2209441-6 0005-2787 nnns volume:606 year:1980 number:2 pages:202-213 http://linkinghub.elsevier.com/retrieve/pii/0005-2787(80)90030-1 GBV_USEFLAG_H ZDB-1-SDJ GBV_NL_ARTICLE AR 606 1980 2 202-213 |
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(DE-627)NLEJ185895921 (DE-599)GBVNLZ185895921 DE-627 ger DE-627 rakwb eng Purification and properties of a DNA ligase from a soluble DNA replication complex 1980 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier A DNA ligase has been purified from a subnuclear soluble replication complex isolated from adenovirus type 2-infected human KB cells. DNA ligase activity could not be demonstrated using an exogenous template until the complex was dissociated, suggesting that the ligase activity may be a component of the complex. The purified enzyme was free of endonuclease, exonuclease, 5'-nucleotidase, and phosphatase activites, and had a molecular weight of 105 000, as estimated by sedimentation in a glycerol gradient. The ligase requires ATP and a divalent cation for activity. The optimum of the reaction is at pH 7.8 in 50-100 mM Tris-HCl buffer and 10-20 mM MgCl"2. Monovalent salts greatly stimulate ligase activity and the optimum was found at 150 mM. The reaction is very sensitive to high temperature; maximum activity was observed at 25-30^oC. ATP is the sole required cofactor and NAD, dATP and GTP could not replace the requirement for ATP. The K"m for ATP is 60 μM. The K"m for DNA is 250 μg/ml or 1.6 nmol of terminal phosphate/ml and thus the enzyme shows relatively weak affinity for exogenous DNA. The maximum conversion of ^3^2P into a phosphatase-resistant form is approximately 1.3% of the total, whereas T4 ligase, under the same conditions, can convert more than 25% of phosphate into a resistant form. Elsevier Journal Backfiles on ScienceDirect 1907 - 2002 Tsuruo, T. oth Arens, M. oth Padmanabhan, R. oth Green, M. oth in BBA Section Nucleic Acids And Protein Synthesis Amsterdam : Elsevier 606(1980), 2, Seite 202-213 (DE-627)NLEJ185870457 (DE-600)2209441-6 0005-2787 nnns volume:606 year:1980 number:2 pages:202-213 http://linkinghub.elsevier.com/retrieve/pii/0005-2787(80)90030-1 GBV_USEFLAG_H ZDB-1-SDJ GBV_NL_ARTICLE AR 606 1980 2 202-213 |
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(DE-627)NLEJ185895921 (DE-599)GBVNLZ185895921 DE-627 ger DE-627 rakwb eng Purification and properties of a DNA ligase from a soluble DNA replication complex 1980 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier A DNA ligase has been purified from a subnuclear soluble replication complex isolated from adenovirus type 2-infected human KB cells. DNA ligase activity could not be demonstrated using an exogenous template until the complex was dissociated, suggesting that the ligase activity may be a component of the complex. The purified enzyme was free of endonuclease, exonuclease, 5'-nucleotidase, and phosphatase activites, and had a molecular weight of 105 000, as estimated by sedimentation in a glycerol gradient. The ligase requires ATP and a divalent cation for activity. The optimum of the reaction is at pH 7.8 in 50-100 mM Tris-HCl buffer and 10-20 mM MgCl"2. Monovalent salts greatly stimulate ligase activity and the optimum was found at 150 mM. The reaction is very sensitive to high temperature; maximum activity was observed at 25-30^oC. ATP is the sole required cofactor and NAD, dATP and GTP could not replace the requirement for ATP. The K"m for ATP is 60 μM. The K"m for DNA is 250 μg/ml or 1.6 nmol of terminal phosphate/ml and thus the enzyme shows relatively weak affinity for exogenous DNA. The maximum conversion of ^3^2P into a phosphatase-resistant form is approximately 1.3% of the total, whereas T4 ligase, under the same conditions, can convert more than 25% of phosphate into a resistant form. Elsevier Journal Backfiles on ScienceDirect 1907 - 2002 Tsuruo, T. oth Arens, M. oth Padmanabhan, R. oth Green, M. oth in BBA Section Nucleic Acids And Protein Synthesis Amsterdam : Elsevier 606(1980), 2, Seite 202-213 (DE-627)NLEJ185870457 (DE-600)2209441-6 0005-2787 nnns volume:606 year:1980 number:2 pages:202-213 http://linkinghub.elsevier.com/retrieve/pii/0005-2787(80)90030-1 GBV_USEFLAG_H ZDB-1-SDJ GBV_NL_ARTICLE AR 606 1980 2 202-213 |
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(DE-627)NLEJ185895921 (DE-599)GBVNLZ185895921 DE-627 ger DE-627 rakwb eng Purification and properties of a DNA ligase from a soluble DNA replication complex 1980 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier A DNA ligase has been purified from a subnuclear soluble replication complex isolated from adenovirus type 2-infected human KB cells. DNA ligase activity could not be demonstrated using an exogenous template until the complex was dissociated, suggesting that the ligase activity may be a component of the complex. The purified enzyme was free of endonuclease, exonuclease, 5'-nucleotidase, and phosphatase activites, and had a molecular weight of 105 000, as estimated by sedimentation in a glycerol gradient. The ligase requires ATP and a divalent cation for activity. The optimum of the reaction is at pH 7.8 in 50-100 mM Tris-HCl buffer and 10-20 mM MgCl"2. Monovalent salts greatly stimulate ligase activity and the optimum was found at 150 mM. The reaction is very sensitive to high temperature; maximum activity was observed at 25-30^oC. ATP is the sole required cofactor and NAD, dATP and GTP could not replace the requirement for ATP. The K"m for ATP is 60 μM. The K"m for DNA is 250 μg/ml or 1.6 nmol of terminal phosphate/ml and thus the enzyme shows relatively weak affinity for exogenous DNA. The maximum conversion of ^3^2P into a phosphatase-resistant form is approximately 1.3% of the total, whereas T4 ligase, under the same conditions, can convert more than 25% of phosphate into a resistant form. Elsevier Journal Backfiles on ScienceDirect 1907 - 2002 Tsuruo, T. oth Arens, M. oth Padmanabhan, R. oth Green, M. oth in BBA Section Nucleic Acids And Protein Synthesis Amsterdam : Elsevier 606(1980), 2, Seite 202-213 (DE-627)NLEJ185870457 (DE-600)2209441-6 0005-2787 nnns volume:606 year:1980 number:2 pages:202-213 http://linkinghub.elsevier.com/retrieve/pii/0005-2787(80)90030-1 GBV_USEFLAG_H ZDB-1-SDJ GBV_NL_ARTICLE AR 606 1980 2 202-213 |
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Purification and properties of a DNA ligase from a soluble DNA replication complex |
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A DNA ligase has been purified from a subnuclear soluble replication complex isolated from adenovirus type 2-infected human KB cells. DNA ligase activity could not be demonstrated using an exogenous template until the complex was dissociated, suggesting that the ligase activity may be a component of the complex. The purified enzyme was free of endonuclease, exonuclease, 5'-nucleotidase, and phosphatase activites, and had a molecular weight of 105 000, as estimated by sedimentation in a glycerol gradient. The ligase requires ATP and a divalent cation for activity. The optimum of the reaction is at pH 7.8 in 50-100 mM Tris-HCl buffer and 10-20 mM MgCl"2. Monovalent salts greatly stimulate ligase activity and the optimum was found at 150 mM. The reaction is very sensitive to high temperature; maximum activity was observed at 25-30^oC. ATP is the sole required cofactor and NAD, dATP and GTP could not replace the requirement for ATP. The K"m for ATP is 60 μM. The K"m for DNA is 250 μg/ml or 1.6 nmol of terminal phosphate/ml and thus the enzyme shows relatively weak affinity for exogenous DNA. The maximum conversion of ^3^2P into a phosphatase-resistant form is approximately 1.3% of the total, whereas T4 ligase, under the same conditions, can convert more than 25% of phosphate into a resistant form. |
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A DNA ligase has been purified from a subnuclear soluble replication complex isolated from adenovirus type 2-infected human KB cells. DNA ligase activity could not be demonstrated using an exogenous template until the complex was dissociated, suggesting that the ligase activity may be a component of the complex. The purified enzyme was free of endonuclease, exonuclease, 5'-nucleotidase, and phosphatase activites, and had a molecular weight of 105 000, as estimated by sedimentation in a glycerol gradient. The ligase requires ATP and a divalent cation for activity. The optimum of the reaction is at pH 7.8 in 50-100 mM Tris-HCl buffer and 10-20 mM MgCl"2. Monovalent salts greatly stimulate ligase activity and the optimum was found at 150 mM. The reaction is very sensitive to high temperature; maximum activity was observed at 25-30^oC. ATP is the sole required cofactor and NAD, dATP and GTP could not replace the requirement for ATP. The K"m for ATP is 60 μM. The K"m for DNA is 250 μg/ml or 1.6 nmol of terminal phosphate/ml and thus the enzyme shows relatively weak affinity for exogenous DNA. The maximum conversion of ^3^2P into a phosphatase-resistant form is approximately 1.3% of the total, whereas T4 ligase, under the same conditions, can convert more than 25% of phosphate into a resistant form. |
| abstract_unstemmed |
A DNA ligase has been purified from a subnuclear soluble replication complex isolated from adenovirus type 2-infected human KB cells. DNA ligase activity could not be demonstrated using an exogenous template until the complex was dissociated, suggesting that the ligase activity may be a component of the complex. The purified enzyme was free of endonuclease, exonuclease, 5'-nucleotidase, and phosphatase activites, and had a molecular weight of 105 000, as estimated by sedimentation in a glycerol gradient. The ligase requires ATP and a divalent cation for activity. The optimum of the reaction is at pH 7.8 in 50-100 mM Tris-HCl buffer and 10-20 mM MgCl"2. Monovalent salts greatly stimulate ligase activity and the optimum was found at 150 mM. The reaction is very sensitive to high temperature; maximum activity was observed at 25-30^oC. ATP is the sole required cofactor and NAD, dATP and GTP could not replace the requirement for ATP. The K"m for ATP is 60 μM. The K"m for DNA is 250 μg/ml or 1.6 nmol of terminal phosphate/ml and thus the enzyme shows relatively weak affinity for exogenous DNA. The maximum conversion of ^3^2P into a phosphatase-resistant form is approximately 1.3% of the total, whereas T4 ligase, under the same conditions, can convert more than 25% of phosphate into a resistant form. |
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<?xml version="1.0" encoding="UTF-8"?><collection xmlns="http://www.loc.gov/MARC21/slim"><record><leader>01000caa a22002652 4500</leader><controlfield tag="001">NLEJ185895921</controlfield><controlfield tag="003">DE-627</controlfield><controlfield tag="005">20230505183848.0</controlfield><controlfield tag="007">cr uuu---uuuuu</controlfield><controlfield tag="008">070506s1980 xx |||||o 00| ||eng c</controlfield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(DE-627)NLEJ185895921</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(DE-599)GBVNLZ185895921</subfield></datafield><datafield tag="040" ind1=" " ind2=" "><subfield code="a">DE-627</subfield><subfield code="b">ger</subfield><subfield code="c">DE-627</subfield><subfield code="e">rakwb</subfield></datafield><datafield tag="041" ind1=" " ind2=" "><subfield code="a">eng</subfield></datafield><datafield tag="245" ind1="1" ind2="0"><subfield code="a">Purification and properties of a DNA ligase from a soluble DNA replication complex</subfield></datafield><datafield tag="264" ind1=" " ind2="1"><subfield code="c">1980</subfield></datafield><datafield tag="336" ind1=" " ind2=" "><subfield code="a">nicht spezifiziert</subfield><subfield code="b">zzz</subfield><subfield code="2">rdacontent</subfield></datafield><datafield tag="337" ind1=" " ind2=" "><subfield code="a">nicht spezifiziert</subfield><subfield code="b">z</subfield><subfield code="2">rdamedia</subfield></datafield><datafield tag="338" ind1=" " ind2=" "><subfield code="a">nicht spezifiziert</subfield><subfield code="b">zu</subfield><subfield code="2">rdacarrier</subfield></datafield><datafield tag="520" ind1=" " ind2=" "><subfield code="a">A DNA ligase has been purified from a subnuclear soluble replication complex isolated from adenovirus type 2-infected human KB cells. DNA ligase activity could not be demonstrated using an exogenous template until the complex was dissociated, suggesting that the ligase activity may be a component of the complex. The purified enzyme was free of endonuclease, exonuclease, 5'-nucleotidase, and phosphatase activites, and had a molecular weight of 105 000, as estimated by sedimentation in a glycerol gradient. The ligase requires ATP and a divalent cation for activity. The optimum of the reaction is at pH 7.8 in 50-100 mM Tris-HCl buffer and 10-20 mM MgCl"2. Monovalent salts greatly stimulate ligase activity and the optimum was found at 150 mM. The reaction is very sensitive to high temperature; maximum activity was observed at 25-30^oC. ATP is the sole required cofactor and NAD, dATP and GTP could not replace the requirement for ATP. The K"m for ATP is 60 μM. The K"m for DNA is 250 μg/ml or 1.6 nmol of terminal phosphate/ml and thus the enzyme shows relatively weak affinity for exogenous DNA. The maximum conversion of ^3^2P into a phosphatase-resistant form is approximately 1.3% of the total, whereas T4 ligase, under the same conditions, can convert more than 25% of phosphate into a resistant form.</subfield></datafield><datafield tag="533" ind1=" " ind2=" "><subfield code="f">Elsevier Journal Backfiles on ScienceDirect 1907 - 2002</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Tsuruo, T.</subfield><subfield code="4">oth</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Arens, M.</subfield><subfield code="4">oth</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Padmanabhan, R.</subfield><subfield code="4">oth</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Green, M.</subfield><subfield code="4">oth</subfield></datafield><datafield tag="773" ind1="0" ind2="8"><subfield code="i">in</subfield><subfield code="t">BBA Section Nucleic Acids And Protein Synthesis</subfield><subfield code="d">Amsterdam : Elsevier</subfield><subfield code="g">606(1980), 2, Seite 202-213</subfield><subfield code="w">(DE-627)NLEJ185870457</subfield><subfield code="w">(DE-600)2209441-6</subfield><subfield code="x">0005-2787</subfield><subfield code="7">nnns</subfield></datafield><datafield tag="773" ind1="1" ind2="8"><subfield code="g">volume:606</subfield><subfield code="g">year:1980</subfield><subfield code="g">number:2</subfield><subfield code="g">pages:202-213</subfield></datafield><datafield tag="856" ind1="4" ind2="0"><subfield code="u">http://linkinghub.elsevier.com/retrieve/pii/0005-2787(80)90030-1</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">GBV_USEFLAG_H</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">ZDB-1-SDJ</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">GBV_NL_ARTICLE</subfield></datafield><datafield tag="951" ind1=" " ind2=" "><subfield code="a">AR</subfield></datafield><datafield tag="952" ind1=" " ind2=" "><subfield code="d">606</subfield><subfield code="j">1980</subfield><subfield code="e">2</subfield><subfield code="h">202-213</subfield></datafield></record></collection>
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7.3997383 |