دورية أكاديمية
Inhibition of [(15)N]valine transamination during selective labeling of barstar in a T7 polymerase system.
| العنوان: | Inhibition of [(15)N]valine transamination during selective labeling of barstar in a T7 polymerase system. |
|---|---|
| المؤلفون: | Lopukhov LV; Kazan Institute of Biochemistry and Biophysics, Kazan Research Center, Russian Academy of Sciences, Kazan 420111, Russia., Ponomareva AA, Yagodina LO |
| المصدر: | Biochemistry. Biokhimiia [Biochemistry (Mosc)] 2004 Jul; Vol. 69 (7), pp. 806-8. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: MAIK Nauka/Interperiodica Country of Publication: United States NLM ID: 0376536 Publication Model: Print Cited Medium: Print ISSN: 0006-2979 (Print) Linking ISSN: 00062979 NLM ISO Abbreviation: Biochemistry (Mosc) Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: <2007->: Moscow : MAIK Nauka/Interperiodica Original Publication: New York, Consultants Bureau [etc.] |
| مواضيع طبية MeSH: | Bacterial Proteins/*metabolism , DNA-Directed RNA Polymerases/*metabolism , Valine/*metabolism , Viral Proteins/*metabolism, Aminooxyacetic Acid/chemistry ; Aminooxyacetic Acid/metabolism ; Bacterial Proteins/chemistry ; Bacterial Proteins/pharmacology ; Catalysis/drug effects ; Escherichia coli/drug effects ; Escherichia coli/genetics ; Escherichia coli/metabolism ; Isopropyl Thiogalactoside/pharmacology ; Magnetic Resonance Spectroscopy ; Nitrogen Isotopes ; Rifampin/pharmacology ; Transaminases/antagonists & inhibitors ; Transaminases/metabolism ; Valine/chemistry |
| مستخلص: | Selective labeling of barstar by the stable (15)N isotope of the valine residue with high selectivity of the label incorporation resulting from the process of gene expression in Escherichia coli BL21(DE3) has been optimized. We have shown that alpha-aminooxyacetic acid (AOAA) significantly reduces the isotope redistribution, thus increasing the selectivity of (15)N incorporation into the synthesized protein, as detected by 2D-NMR. Quantitative measurements were used to determine the selectivity for the incorporation of isotope-labeled valine residue, which was 96% in the case using AOAA. Studies of the dynamics of barstar synthesis showed that no suppression of barstar yield is observed under the regulation of the T7 polymerase expression system by isopropylthio-beta-D-galactoside (IPTG) and rifampicin using AOAA. |
| المشرفين على المادة: | 0 (Bacterial Proteins) 0 (Nitrogen Isotopes) 0 (Viral Proteins) 14I68GI3OQ (Aminooxyacetic Acid) 367-93-1 (Isopropyl Thiogalactoside) 37328-61-3 (barstar protein, Bacillus amyloliquefaciens) EC 2.6.1.- (Transaminases) EC 2.7.7.- (bacteriophage T7 RNA polymerase) EC 2.7.7.6 (DNA-Directed RNA Polymerases) HG18B9YRS7 (Valine) VJT6J7R4TR (Rifampin) |
| تواريخ الأحداث: | Date Created: 20040818 Date Completed: 20080721 Latest Revision: 20190917 |
| رمز التحديث: | 20221213 |
| DOI: | 10.1023/b:biry.0000040207.23339.21 |
| PMID: | 15310282 |
| قاعدة البيانات: | MEDLINE |
Find this article in full text from Springer Verlag. 
Full Text Finder | تدمد: | 0006-2979 |
|---|---|
| DOI: | 10.1023/b:biry.0000040207.23339.21 |