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Determination of Mammillaria gracillis N-glycan Patterns by ESI Q-TOF Mass Spectrometry

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Title Determination of Mammillaria gracillis N-glycan Patterns by ESI Q-TOF Mass Spectrometry
Određivanje N-glycana u uzorcima tkiva kaktusa Mammillaria gracillis masenom spektrometrijom ESI Q-TOF
 
Creator Balen, Biljana
Zamfir, Alina
Vakhrushev, Sergey Y.
Krsnik-Rasol, Marijana
Peter-Katalinić, Jasna
 
Subject Cactaceae; crown-gall tumour; morphogenesis; N-glycans; sialic acid
 
Description N-glycans detached from the 42 kDa cellular glycoprotein of in vitro propagated cactus (Mammillaria gracillis Pfeiff.) shoots, callus, hyperhydric regenerants and TW tumour were analyzed by mass spectrometry after in-gel deglycosylation of excised protein bands. Mixtures of N-glycans were shown by the electrospray MS analysis to be highly complex and heterogeneous. Only a low amount of high-mannose N-glycans was present, while the majority of structures were of the complex-type N-glycans. A single N-glycan structure was observed in the shoot sample, which appeared to be in good correlation with previous results. Similarities of assigned structures were revealed in the callus and tumour tissue while the highest number of different oligosaccharide structures was found in the hyperhydric regenerant. The presence of sialic acid as a constituent of N-glycans was indicated by lectin staining, ion exchange chromatography and molecular mapping by mass spectrometry, and could be correlated with the differentiation status of the tissues. Results obtained in this study indicate that different morphological levels can be correlated with the N-glycan composition of cellular glycoproteins.
N-glikani, oslobođeni sa staničnoga glikoproteina od 42 kDa iz izdanka, kalusa, hiperhidriranoga regeneranta i tumora TW uzgojenih u kulturi tkiva kaktusa Mammillaria gracillis Pfeiff. analizirani su masenom spektrometrijom nakon deglikozilacije u gelu. Analiza masenom spektrometrijom ESI pokazala je kako su smjese oslobođ enih oligosaharida vrlo složene i heterogene, a većina struktura pripadala je kompleksnome tipu N-glikana. U uzorku izdanka opažena je samo jedna struktura, {to je u skladu s prija{njim rezultatima. Kalusno i tumorsko tkivo pokazalo je mnogo sličnosti u pripisanim strukturama dok je najveći broj različitih oligosaharida prona- đen u hiperhidriranome regenerantu. Na prisutnost sijalinske kiseline ukazala je inkubacija s lektinima, kromatografija ionske izmjene te masena spektrometrija. Rezultati dobiveni u ovim istraživanjima pokazuju da se stupanj organiziranosti tkiva može povezati sa sastavom N-glikana staničnih glikoproteina.
 
Publisher Croatian Chemical Society
 
Date 2005-09-15
 
Type text
info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
 
Format application/pdf
 
Identifier http://hrcak.srce.hr/58
http://hrcak.srce.hr/file/58
 
Source Croatica Chemica Acta; Vol.78 No.3; ISSN 0011-1643 (Print); ISSN 1334-417X (Online)
 
Language en
 
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Parts of the contents of Croat. Chem. Acta (e. g. figures or tables) may be reproduced without prior permission, provided reference is made to their source.