Bridging Reagent for Protein

Publisher Website
Google Scholar
Abstract
1. Several N, N'-polymethylene-bis (iodoacetamide) were synthesized and their reaction with cysteine was investigated. 2. Rabbit muscle aldolase [EC 4.1.2.13] was treated with N, N'-polymethylene-bis (iodoacetamide). When a high concentration of the enzyme and an enough amount of the brigding reagent were used, insoluble protein with an appreciable enzymatic activity was obtained in a yield of 50%. 3.When the reaction with aldolase was carried out using a dilute enzyme solution and limited amounts of bridging reagents, all the enzyme protein remained soluble. The sedimentation coefficient of the major peak was the same as that of native aldolase at pH 7.5, showing that the extent of inter-molecular bridging reaction was small. The content of S-carboxymethylcysteine of the preparation was about 6 moles per mole of aldolase. The sedimentation coefficients of the acid dissociated aldorase and of the bridged aldolase were 1.6S and 2.3–2.5S, respectively. Therefore, an inter-subunit bridging must have occurred during the reaction. The bridged aldolase preparation showed no tendency of stabilization towards acid, alkali and urea denaturation. Acid denatured bridged aldorase recover less enzymatic activity on neutralization than that in the case of aldolase.

This publication has 0 references indexed in Scilit: