New devices for flow dialysis and ultrafiltration for the study of protein-ligand interactions

✍ Scribed by Knut Feldmann

• Book ID: 102985586
• Publisher: Elsevier Science
• Year: 1978
• Tongue: English
• Weight: 643 KB
• Volume: 88
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(78)90414-1

✦ Synopsis

A flow dialysis procedure with an approach to steady state in less than half a minute has been developed. The main advantage of the new device is that it permits the use of lower levels of radioactivity and therefore allows determination of dissociation constants (K,) of tight complexes, i.e., with K, values of 10-r M or even lower. An ultrafiltration device was also developed which does not disturb binding equilibria because it uses only a small fraction of the ultrafiltrate (about 1% of the total solution) and which allows simultaneous measurements of six samples in a few minutes. Both methods were tested with rabbit skeletal muscle glycogen phosphorylases b and a (EC 2.4.1.1) and the allosteric ligand 5'-AMP. The K, values are in good agreement with those previously obtained by conventional methods.