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  • Title: Analysis of hydroxyproline and hydroxyproline-arabinosides of plant origin by high-performance anion-exchange chromatography-pulsed amperometric detection.
    Author: Campargue C, Lafitte C, Esquerré-Tugayé MT, Mazau D.
    Journal: Anal Biochem; 1998 Mar 01; 257(1):20-5. PubMed ID: 9512767.
    Abstract:
    The plant cell wall hydroxyproline-rich glycoprotein (HRGP), also called extensin, contains arabinose and oligoarabinoside 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(OH)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.
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