Affinity Chromatographic Purification and Characterization of Sialic Acid Binding Proteins in Bovine Kidney
書誌事項
- タイトル別名
-
- Affinity Chromatographic Purification a
この論文をさがす
説明
application/pdf
紀要論文
Sialic acid binding proteins were purified from bovine kidney by successive affinity chromatography on fetuin and heparin clumns. The proteins adsorbed calcium-dependently on fetuin-Sepharose were further subjected to affinity chromatography on heparin-Sepharose. The proteins were separated into two fractions, fraction I and fraction II, by elution with 2mM EDTA and 0.3M NaCl, respectively. The affinity purified fractions had the binding activities to biotinylated fetuin in a polystyrene microtiter well, but not hemagglutinating activities. Inhibition assay of the binding revealed that N-acetylneuraminic acid is the most potent inhibitor for both fractions among the monosaccharides tested. Upon SDS-polyacrylamide gel electrophoresis, fraction-I gave three bands corresponding to 37kDa, 43kDa and 50kDa proteins and fraction-II four bands corresponding to 38kDa, 39kDa, 41kDa and 74kDa proteins. The proteins were electroblotted onto a polyvinylidene difluoride membrane and then subjected to the direct chemical analyses and the binding studies using horseradish peroxidase (HRP)-labeled binding probes. All the proteins had\ similar amino acid compositions and were N-terminally blocked. Neither of all proteins was stained with HRP-concanavalin A and HRP-peanut agglutinin, suggesting that they are not glycosylated. All of them were stained with HRP-fetuin and HRP-anhydrotrypsin. The results suggest that they have the carbohydrate recognition domain specific to N-acetylneuraminic acid and are the fragments produced by proteolytic digestion with endogenous trypsin family proteases in the kidney.
収録刊行物
-
- お茶の水女子大學自然科學報告
-
お茶の水女子大學自然科學報告 42 (1), 1-11, 1991-07-01
お茶の水女子大学
- Tweet
詳細情報 詳細情報について
-
- CRID
- 1050282677926124032
-
- NII論文ID
- 110005944153
-
- NII書誌ID
- AN00033958
-
- ISSN
- 00298190
-
- HANDLE
- 10083/808
-
- NDL書誌ID
- 3733892
-
- 本文言語コード
- en
-
- 資料種別
- departmental bulletin paper
-
- データソース種別
-
- IRDB
- NDL
- CiNii Articles