Immobilized carboxypeptidase Y. applications in protein chemistry

Cross‐linking of reversibly immobilized enzymes

“…HMD-CL-Sepharose was prepared by the method of Liberatore et al [2] with modifications to reduce the amount of bound HMD. For this study the washed gel was oxidized for IO min at room temperature with 0.2 mM NaI04.…”

“…CL-Sepharose-glutathione was prepared by the method of Liberatore et al [2] ; it was reacted with 2,2'-dipyridyl disulfide and papain according to Brocklehurst, et al [4]. The bound enzyme derivative contained 1 mg protein ml/gel.…”

“…The procedure of Weber and Osborn [ 131 was used with the modification that mercaptoethanol was not added to protein solutions (l), (2) and (3) to prevent chain separation of cY-trypsin. substantial activity against BAEE and essentially complete activity against N,N'-dimethyl bovine serum albumin.…”

Immobilization of invertase on sepharose-linked enzyme glycosyl recognizing polyclonal antibodies

“…The recalcitrance of glycoenzymes to covalent immobilization procedures involving amino acid side chain groups (Liberatore et al, 1976;Pollock et al, 1978) necessitated strategies in which the glycosyl residues will form the point of attachment between the enzyme and the support (Hsiao and Royer, 1979;Woodward and Wiseman, 1978). Among these, the affinity immobilization procedure employing the lectin concanavalin A (con A) appears to be the most promising (Saleemuddin and Husain, 1991).…”

Mechanistic studies of carboxypeptidase Y from Saccharomyces cerevisiae. pH and pD profiles and inactivation at low pH (pD) values

“…Carboxypeptidase Y is a relatively non-specific single-chain exopeptidase with no metalion-dependence (Hayashi et al, 1973 and no intrinsic endopeptidase activity (Hermodsen et al, 1972;Hayashi et al, 1973;Lee & Riordan, 1978), capable of cleaving even proline residues from proteins. [tshigh thermal stability, apparent resistance to pH extrema and activity in 6M-urea or in sodium dodecyl sulphate media have led to its increasing use in protein sequencing and structural studies (Hayashi, 1977;Liberatore et al, 1976;Martin et al, 1977).…”