Academic Journal
Purification of Methionyl‐tRNA Synthetase from Escherichia coli K12 by Affinity Chromatography
| العنوان: | Purification of Methionyl‐tRNA Synthetase from Escherichia coli K12 by Affinity Chromatography |
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| المؤلفون: | Robert‐Gero, Malka, Waller, Jean‐Pierre |
| المصدر: | European Journal of Biochemistry ; volume 31, issue 2, page 315-319 ; ISSN 0014-2956 1432-1033 |
| بيانات النشر: | Wiley |
| سنة النشر: | 1972 |
| المجموعة: | Wiley Online Library (Open Access Articles via Crossref) |
| الوصف: | An affinity column for the purification of methionyl‐tRNA synthetase has been prepared. Cyanogen‐bromide‐activated Sepharose 4B was reacted successively with hexamethylene‐diamine and N ‐nitrophenylsulphenylmethionine succinimide ester. The nitrophenylsulphenyl protecting group of the polymer was removed under mild conditions with aqueous sodium thiosulfate. The affinity columns were equilibrated in 200 mM potassium phosphate buffer; this ionic strength proved to be sufficiently high to avoid the retention of methionyl‐tRNA synthetase by ion exchange. On columns with high methionine concentration (8–20 μmol/ml gel bed), the enzyme was strongly retained but could be selectively eluted by addition of 20 mM methionine to the buffer. However this resulted in excessive dilution of the enzyme. Highly purified enzyme could be obtained in high yield by lowering the methionine substitution of the polymer to 4.5 μmol/ml gel and by introducing the affinity chromatography after the first DEAE‐Sephadex step of the conventional purification procedure. Under these conditions no retention but sufficient retardation of the enzyme on the column was observed. The possibility of generalizing the method for the purification of other proteins recognizing an amino acid with its free amino group is discussed. |
| نوع الوثيقة: | article in journal/newspaper |
| اللغة: | English |
| DOI: | 10.1111/j.1432-1033.1972.tb02535.x |
| الاتاحة: | http://dx.doi.org/10.1111/j.1432-1033.1972.tb02535.x https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1111%2Fj.1432-1033.1972.tb02535.x https://febs.onlinelibrary.wiley.com/doi/pdf/10.1111/j.1432-1033.1972.tb02535.x |
| Rights: | http://onlinelibrary.wiley.com/termsAndConditions#vor |
| رقم الانضمام: | edsbas.D981E8BD |
| قاعدة البيانات: | BASE |
| DOI: | 10.1111/j.1432-1033.1972.tb02535.x |
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