Actin immobilization on chitin for purifying myosin II

• Published in: Biochemistry and molecular biology education 2008-01, Vol.36 (1), p.55-60
• Main Authors: de Souza, Marcelle Gomes, Grossi, André Luiz, Pereira, Elisângela Lima Bastos, da Cruz, Carolina Oliveira, Mendes, Fernanda Machado, Cameron, Luiz Claudio, Paiva, Carmen Lucia Antão
• Format: Article
• Language: English
• Subjects: actin immobilization; affinity chromatography; molecular cell biology; Myosin purification; teaching protein chemistry
• ISSN: 1470-8175; 1539-3429
• DOI: 10.1002/bmb.122

This article presents our experience on teaching biochemical sciences through an innovative approach that integrates concepts of molecular cell biology and protein chemistry. This original laboratory exercise is based on the preparation of an affinity chromatography column containing F‐actin molecules immobilized on chitin particles for purifying skeletal myosin II. It favors the active learning of protein extraction and purification, the learning of concepts such as muscle contraction, cytoskeleton structure, and its importance for the living cell. This laboratory exercise also promotes learning biotechnological applications of chitin and the applications of protein immobilization in different industrial fields. Furthermore, the activities target the development of laboratorial abilities, problem‐solving skills, and the ability to write a scientific report, following the model of a scientific article. The trials are mainly proposed for either an undergraduate project for advanced students in the life sciences or a postgraduate practical training course. In both the cases, the students must have had biochemistry as part of their regular curriculum. Alternatively, the affinity chromatography method can fit in any regular biochemistry course if active chitin, F‐actin, and a myosin II extract are provided. It is very important to mention that this laboratory exercise can be used even in places where a facility such as ultracentrifugation is lacking. For that, the steps of actin purification are skipped, and actin is commercially obtained. Therefore, it is an adequate approach for the active learning of biochemical and molecular cell biology principles and techniques even in poor countries.