Detection of aldolase activity on polyacrylamide gels: Application to 2-keto-4-hydroxyglutarate aldolase

✍ Scribed by Neil D. Lewinski; Eugene E. Dekker

• Book ID: 102982902
• Publisher: Elsevier Science
• Year: 1978
• Tongue: English
• Weight: 935 KB
• Volume: 87
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(78)90568-7

✦ Synopsis

A method is described for the detection of 2-keto-4-hydroxyglutarate aldolase activity after electrophoresis of the enzyme on polyacrylamide gels. When gels are incubated with substrate (2-keto-4-hydroxyglutarate), activity is seen as a yellow-colored band due to interaction' of the product (glyoxylate) with ortho-aminobenzaldehyde and glycine. Positive results have been obtained using either crude cell-free preparations or homogeneous enzyme from Escherichia coli as well as with highly purified samples of aldolase from bovine liver or kidney extracts. The method is potentially applicable to other aldolases that liberate an aliphatic aldehyde as a product; modifications and limitations of the procedure for detecting fructose 1.6-diphosphate aldolase, 2-keto-3-deoxy-6phosphogluconate aldolase, and 2-deoxyribose-5-phosphate aldolase activities have been explored.