Analysis of hydroxyproline and hydroxyproline-arabinosides of plant origin by high-performance anion-exchange chromatography pulsed amperometric detection

被引:14
作者
Campargue, C [1 ]
Lafitte, C [1 ]
Esquerré-Tugayé, MT [1 ]
Mazau, D [1 ]
机构
[1] Univ Toulouse 3, UMR CNRS, UPS 5546, Ctr Biol & Physiol Vegetales, F-31062 Toulouse 04, France
关键词
D O I
10.1006/abio.1997.2526
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The plant cell wall hydroxyproline-rich glycoprotein (HRGP), also called extensin, contains arabinose and oligoarabinaside side chains O-glycosidically linked to hydroxyproline (Hyp). We present a highly sensitive method for determining both the glycosylation pattern and the Hyp content of HRGP requiring only nanomole amounts of each Hyp-compound for accurate determination. This method is based on anion-exchange chromatography followed by pulsed amperometric detection of the Hyp-oligoarabinosides and Hyp released from HRGP by 0.22 M Ba(ON)(2) hydrolysis, which cleaves only peptidyl bonds. A sodium acetate gradient. (0-250 mM) in 150 mM NaOH elutes Hyp and the Hyp-oligoarabinosides Hyp-(Ara)(1-5) in less than 40 min. We have used this procedure to determine the glycosylation pattern of Hyp in plant cell walls, without prior isolation of HRGP. (C) 1998 Academic Press.
引用
收藏
页码:20 / 25
页数:6
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