2024-03-29T11:43:40Z
https://teapot.lib.ocha.ac.jp/oai
oai:teapot.lib.ocha.ac.jp:00034739
2022-12-12T05:39:14Z
347:359:654
Affinity Chromatographic Purification and Characterization of Sialic Acid Binding Proteins in Bovine Kidney
Kojima, Kyoko
Adachi, Masami
Ogawa, Haruko
Seno, Nobuko
Matsumoto, Isamu
400
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.
お茶の水女子大学
1991-07-01
eng
departmental bulletin paper
http://hdl.handle.net/10083/808
https://teapot.lib.ocha.ac.jp/records/34739
AN00033958
00298190
お茶の水女子大學自然科學報告
42
1
1
11
https://teapot.lib.ocha.ac.jp/record/34739/files/KJ00004470757.pdf
application/pdf
933.6 kB
2018-04-19