Highly purified recombinant varicella Zoster virus thymidine kinase is a homodimer
Copyright 2000 Academic Press..
Recombinant varicella zoster virus (VZV) thymidine kinase (TK) was isolated in a fast and gentle two-step procedure from Escherichia coli. The TK was expressed as a PreScission-cleavable fusion protein and purified by glutathione and ATP affinity chromatography, yielding homogeneous, highly pure VZV TK. The purified enzyme displays enzymatic activities with K(m) values of 0.3 +/- 0.06 microM for the natural substrate thymidine and 11.6 +/- 3.2 microM for ATP, indicating the biochemical equivalence with the viral VZV TK expressed in infected cells. Determinations of the native molecular weight by size exclusion chromatography and native polyacrylamide gel electrophoresis revealed that the pure enzyme is biologically active as a homodimer.
| Medienart: |
Artikel |
|---|
| Erscheinungsjahr: |
2000 |
|---|---|
| Erschienen: |
2000 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:18 |
|---|---|
| Enthalten in: |
Protein expression and purification - 18(2000), 3 vom: 01. Apr., Seite 338-45 |
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Amrhein, I [VerfasserIn] |
|---|
| Themen: |
EC 2.7.1.21 |
|---|
| Anmerkungen: |
Date Completed 07.06.2000 Date Revised 15.11.2006 published: Print Citation Status MEDLINE |
|---|
| Förderinstitution / Projekttitel: |
|
|---|
| PPN (Katalog-ID): |
NLM106611763 |
|---|
| LEADER | 01000naa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLM106611763 | ||
| 003 | DE-627 | ||
| 005 | 20231222141231.0 | ||
| 007 | tu | ||
| 008 | 231222s2000 xx ||||| 00| ||eng c | ||
| 028 | 5 | 2 | |a pubmed24n0356.xml |
| 035 | |a (DE-627)NLM106611763 | ||
| 035 | |a (NLM)10733888 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Amrhein, I |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Highly purified recombinant varicella Zoster virus thymidine kinase is a homodimer |
| 264 | 1 | |c 2000 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ohne Hilfsmittel zu benutzen |b n |2 rdamedia | ||
| 338 | |a Band |b nc |2 rdacarrier | ||
| 500 | |a Date Completed 07.06.2000 | ||
| 500 | |a Date Revised 15.11.2006 | ||
| 500 | |a published: Print | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Copyright 2000 Academic Press. | ||
| 520 | |a Recombinant varicella zoster virus (VZV) thymidine kinase (TK) was isolated in a fast and gentle two-step procedure from Escherichia coli. The TK was expressed as a PreScission-cleavable fusion protein and purified by glutathione and ATP affinity chromatography, yielding homogeneous, highly pure VZV TK. The purified enzyme displays enzymatic activities with K(m) values of 0.3 +/- 0.06 microM for the natural substrate thymidine and 11.6 +/- 3.2 microM for ATP, indicating the biochemical equivalence with the viral VZV TK expressed in infected cells. Determinations of the native molecular weight by size exclusion chromatography and native polyacrylamide gel electrophoresis revealed that the pure enzyme is biologically active as a homodimer | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 7 | |a Recombinant Fusion Proteins |2 NLM | |
| 650 | 7 | |a Thymidine Kinase |2 NLM | |
| 650 | 7 | |a EC 2.7.1.21 |2 NLM | |
| 700 | 1 | |a Wurth, C |e verfasserin |4 aut | |
| 700 | 1 | |a Bohner, T |e verfasserin |4 aut | |
| 700 | 1 | |a Hofbauer, R |e verfasserin |4 aut | |
| 700 | 1 | |a Folkers, G |e verfasserin |4 aut | |
| 700 | 1 | |a Scapozza, L |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t Protein expression and purification |d 1991 |g 18(2000), 3 vom: 01. Apr., Seite 338-45 |w (DE-627)NLM012635359 |x 1096-0279 |7 nnns |
| 773 | 1 | 8 | |g volume:18 |g year:2000 |g number:3 |g day:01 |g month:04 |g pages:338-45 |
| 912 | |a GBV_USEFLAG_A | ||
| 912 | |a GBV_NLM | ||
| 951 | |a AR | ||
| 952 | |d 18 |j 2000 |e 3 |b 01 |c 04 |h 338-45 | ||