Non-specific protein binding by adsorbents designed for the specific affinity chromatography of sialidase in crude bacterial extracts

A number of derivatives of Sepharose 4B gels were prepared containing tyramine, -Gly-Gly-Tyr and the tetrapeptide, Thr (But)-Phe-Pro-Tyr-. A specific inhibitor of sialidase, p-diazo-phenyl oxamic acid, was also coupled to these phenol-substituted Sepharose gels. The effectiveness of these gels to purify sialidase by "affinity" chromatography was examined.The sialidase from Vibrio cholerae was adsorbed by these gels and subsequently re-eluted by a change in pH and temperature. The adsorption and elution of enzyme was independent of the presence or absence of the "specific inhibitor" of the sialidase, phenyl oxamic acid. A mechanism for the adsorption and subsequent elution of the sialidase is proposed.It is important to re-evaluate the biological data obtained with sialidase prepared by the affinity column recommended by Cuatrecasas and Illiano, Biochem. Biophys. Res. Commun. 44, 178-184 (1971), since glycosidases from Clostridium perfringens other than sialidase are also adsorbed and eluted under the conditions specified.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/22211/1/0000644.pd