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  • Immunoaffinity purification of the class V chitin synthase of Wangiella (Exophiala) dermatitidis.

Immunoaffinity purification of the class V chitin synthase of Wangiella (Exophiala) dermatitidis.

Preparative biochemistry & biotechnology (2009-05-12)
Dariusz Abramczyk, Paul J Szaniszlo
ABSTRACT

The class V chitin synthase is unique because it has a myosin motor-like domain fused to its catalytic domain. The biochemical properties of this enzyme and its function remain undefined beyond the knowledge that it is the only single chitin synthase required for sustained cell growth at elevated temperatures and, consequently, virulence. This report describes our successful efforts to isolate and purify an active and soluble form of the enzyme from the cell membranes of Wangiella by using a specific polyclonal antibody. To our knowledge, this is the first purification of a single chitin synthase of a filamentous fungus.

MATERIALS
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Product Description

Sigma-Aldrich
NBT-BCIP® solution, BioReagent, suitable as substrate for alkaline phosphatase in dot blots