E-Book Overview
Renato Iozzo and an authoritative team of investigators present for the first time a comprehensive and up-to-date collection of readily reproducible preparative and analytical methods for the in-depth analysis of these important compounds. Featuring step-by-step protocols, this book will enable both novice and experienced researchers to isolate intact proteoglycans from tissues and cultured cells, to establish the composition of their carbohydrate moieties, and to generate strategies for prokaryotic and eukaryotic expression. There are also detailed techniques for the suppression of specific proteoglycan genes, for the detection of mutant cells and their degradation products, and for studying specific interactions between proteoglycans and extracellular matrix proteins and with growth factors and their receptors.
E-Book Content
Methods in Molecular Biology
TM
VOLUME 171
Proteoglycan Protocols Edited by
Renato V. Iozzo, MD
HUMANA PRESS
Isolation from Cell Cultures and Tissues
3
1 Isolation of Proteoglycans from Cell Cultures and Tissues Masaki Yanagishita
1. Introduction Proteoglycans are a class of glycosylated proteins characterized by the presence of glycosaminoglycans as a carbohydrate component, which endow them with unique biological as well as biochemical properties. Therefore, isolation of proteoglycans from various biological sources such as cell cultures and tissues could be achieved by ordinary molecular purification procedures utilizing their general molecular properties and by those taking advantages of the presence of glycosaminoglycan moiety. This chapter focuses on the latter experimental procedures, which are particularly useful for obtaining total proteoglycan and glycosaminoglycan species from various biological sources. These protocols could be followed by purification proced