Detail
Článek
Článek online
FT
Medvik - BMČ
  • Je něco špatně v tomto záznamu ?

Analysis of the glycosylation pattern of plant copper amine oxidases by MALDI-TOF/TOF MS coupled to a manual chromatographic separation of glycans and glycopeptides

V. Franc, P. Řehulka, R. Medda, A. Padiglia, G. Floris, M. Šebela,

. 2013 ; 34 (16) : 2357-67.

Jazyk angličtina Země Německo

Typ dokumentu časopisecké články, práce podpořená grantem

Perzistentní odkaz   https://www.medvik.cz/link/bmc14051127

The N-glycosylation in pea seedling amine oxidase and lentil seedling amine oxidase was analyzed in the present work. For that purpose, the enzymes were purified as native proteins from their natural sources. An enzymatic deglycosylation of pea seedling amine oxidase by endoglycosidase H under denaturing conditions combined with its proteolytic digestion by trypsin was carried out in order to analyze both N-glycans and "trimmed" N-glycopeptides with a residual N-acetylglucosamine attached at the originally occupied N-glycosylation sites. The released N-glycans were subjected to a manual chromatographic purification followed by MALDI-TOF/TOF MS. MS and MS/MS analyses were also performed directly on peptides and N-glycopeptides generated by proteolytic digestion of the studied enzymes. Sequencing of glycopeptides by MALDI-TOF/TOF MS/MS after their separation on a RP using a microgradient chromatographic device clearly demonstrated binding of paucimannose and hybrid N-glycan structures at Asn558. Such carbohydrates have been reported to exist in many plant N-glycoproteins, e.g. in peroxidases. Although high-mannose glycan structures were identified after the enzymatic deglycosylation, they could not be assigned to a particular N-glycosylation site. The presence of unoccupied glycosylation sites in several peptides was also confirmed from MS/MS results.

Citace poskytuje Crossref.org

000      
00000naa a2200000 a 4500
001      
bmc14051127
003      
CZ-PrNML
005      
20140402111752.0
007      
ta
008      
140401s2013 gw f 000 0|eng||
009      
AR
024    7_
$a 10.1002/elps.201200622 $2 doi
035    __
$a (PubMed)23580492
040    __
$a ABA008 $b cze $d ABA008 $e AACR2
041    0_
$a eng
044    __
$a gw
100    1_
$a Franc, Vojtěch
245    10
$a Analysis of the glycosylation pattern of plant copper amine oxidases by MALDI-TOF/TOF MS coupled to a manual chromatographic separation of glycans and glycopeptides / $c V. Franc, P. Řehulka, R. Medda, A. Padiglia, G. Floris, M. Šebela,
520    9_
$a The N-glycosylation in pea seedling amine oxidase and lentil seedling amine oxidase was analyzed in the present work. For that purpose, the enzymes were purified as native proteins from their natural sources. An enzymatic deglycosylation of pea seedling amine oxidase by endoglycosidase H under denaturing conditions combined with its proteolytic digestion by trypsin was carried out in order to analyze both N-glycans and "trimmed" N-glycopeptides with a residual N-acetylglucosamine attached at the originally occupied N-glycosylation sites. The released N-glycans were subjected to a manual chromatographic purification followed by MALDI-TOF/TOF MS. MS and MS/MS analyses were also performed directly on peptides and N-glycopeptides generated by proteolytic digestion of the studied enzymes. Sequencing of glycopeptides by MALDI-TOF/TOF MS/MS after their separation on a RP using a microgradient chromatographic device clearly demonstrated binding of paucimannose and hybrid N-glycan structures at Asn558. Such carbohydrates have been reported to exist in many plant N-glycoproteins, e.g. in peroxidases. Although high-mannose glycan structures were identified after the enzymatic deglycosylation, they could not be assigned to a particular N-glycosylation site. The presence of unoccupied glycosylation sites in several peptides was also confirmed from MS/MS results.
650    _2
$a histaminasa $x analýza $x chemie $x metabolismus $7 D006631
650    _2
$a sekvence aminokyselin $7 D000595
650    _2
$a glykopeptidy $x analýza $x chemie $x izolace a purifikace $7 D006020
650    _2
$a glykosylace $7 D006031
650    _2
$a Lathyrus $x chemie $x enzymologie $7 D027807
650    _2
$a mannosyl-glykoprotein endo-beta-N-acetylglukosaminidasa $x chemie $7 D017038
650    _2
$a molekulární modely $7 D008958
650    _2
$a molekulární sekvence - údaje $7 D008969
650    _2
$a rostlinné proteiny $x analýza $x chemie $x metabolismus $7 D010940
650    _2
$a polysacharidy $x analýza $x chemie $x izolace a purifikace $7 D011134
650    _2
$a sekvenční seřazení $7 D016415
650    _2
$a spektrometrie hmotnostní - ionizace laserem za účasti matrice $x metody $7 D019032
655    _2
$a časopisecké články $7 D016428
655    _2
$a práce podpořená grantem $7 D013485
700    1_
$a Řehulka, Pavel $u -
700    1_
$a Medda, Rosaria $u -
700    1_
$a Padiglia, Alessandra $u -
700    1_
$a Floris, Giovanni $u -
700    1_
$a Šebela, Marek $u -
773    0_
$w MED00001508 $t Electrophoresis $x 1522-2683 $g Roč. 34, č. 16 (2013), s. 2357-67
856    41
$u https://pubmed.ncbi.nlm.nih.gov/23580492 $y Pubmed
910    __
$a ABA008 $b sig $c sign $y a $z 0
990    __
$a 20140401 $b ABA008
991    __
$a 20140402111832 $b ABA008
999    __
$a ok $b bmc $g 1018263 $s 849707
BAS    __
$a 3
BAS    __
$a PreBMC
BMC    __
$a 2013 $b 34 $c 16 $d 2357-67 $i 1522-2683 $m Electrophoresis $n Electrophoresis $x MED00001508
LZP    __
$a Pubmed-20140401

Najít záznam

Citační ukazatele

Nahrávání dat ...

Možnosti archivace

Nahrávání dat ...