Identification of Glycosaminoglycans Using High-Performance Liquid Chromatography on a Hydroxyapatite Column

✍ Scribed by Hozumi Narita; Yusuke Takeda; Keiichi Takagaki; Toshiya Nakamura; Seiko Harata; Masahiko Endo

• Publisher: Elsevier Science
• Year: 1995
• Tongue: English
• Weight: 67 KB
• Volume: 232
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1006/abio.1995.9949

✦ Synopsis

Recently, Takagaki et al. developed a new method

Glycosaminoglycans (heparin, heparan sulfate, derfor the isolation of GAGs with endo-b-xylosidase (5) matan sulfate, chondroitin sulfate, and hyaluronic and fluoro-labeling with 2-aminopyridine (PA). Picoacid) were labeled with a fluorescent reagent, 2-aminomole quantities of PA-GAGs were separated and quanpyridine. The fluoro-labeled glycosaminoglycans were tified using high-performance liquid chromatography subjected to high-performance liquid chromatography (HPLC) with TSKgel SAX (6). This method, however, on a hydroxyapatite column. The binding property of had to be accompanied by digestion with chondroitineach glycosaminoglycan to hydroxyapatite was differases to identify dermatan sulfate and chondroitin sulent. The structural properties of glycosaminoglycans fate.

bound to hydroxyapatite were then investigated using

In this paper, we describe a procedure for identifying chemical desulfated or enzymic depolymerized glycos-PA-GAGs using HPLC on a hydroxyapatite column. aminoglycans. This revealed that the sulfate content We further report that the separation of the GAGs, and molecular weight of the glycosaminoglycans corparticularly dermatan sulfate and chondroitin sulfate, related with their binding properties to hydroxyapawere based on their content of iduronic acid residues.

tite. Desulfated dermatan sulfate but not desulfated chondroitin 6-sulfate bound to the hydroxyapatite.

MATERIALS AND METHODS These data indicate that iduronic acid residues of glycosaminoglycans are important for the binding prop-2 Abbreviations used: GAG, glycosaminoglycan; PA, 2-aminopyridine; HPLC, high-performance liquid chromatography. 10 3 , and 5.0 1 10 3 ) were purchased from Sigma Chemi-133