title: N-glycosylation patterns in two α-l-arabinofuranosidases from Penicillium canescens belonging to the glycoside hydrolase families 51 and 54.
abstract: Using MALDI-TOF mass spectrometry (MS) peptide fingerprinting procedure followed by the analysis of MS data with the GlycoMod tool from the ExPASy proteomic site, N-glycosylation of two GH51 and GH54 family α-l-arabinofuranosidases (Abf51A and Abf54A) from Penicillium canescens was studied. Variable N-linked glycans were identified at five out of eight potential N-glycosylation sites in the Abf51A and one out of three potential N-glycosylation sites in the Abf54A. The discriminated glycans represented high-mannose oligosaccharides (Man)x(GlcNAc)2 with a number of Man residues up to 7 or the products of sequential enzymatic trimming of a high-mannose glycan with α-mannosidases and β-N-acetylhexosaminidases. The Abf54A peptide, containing the Asn254 glycosylation site, and one peptide from the Abf51A, containing the Asn163 glycosylation site, were found to exist not only in glycosylated, but also in a native non-modified form.
authors.affiliation: Department of Chemistry, M.V. Lomonosov Moscow State University, Vorobyovy Gory 1/11, Moscow 119991, Russia; A.N. Bach Institute of Biochemistry, Russian Academy of Sciences, Leninsky Pr. 33, Moscow 119071, Russia. Electronic address: avgusakov@enzyme.chem.msu.ru.
authors.forename: Alexander V
authors.initials: AV
authors.lastname: Gusakov
authors.forename: Olga A
authors.initials: OA
authors.lastname: Sinitsyna
authors.forename: Alexandra M
authors.initials: AM
authors.lastname: Rozhkova
authors.forename: Arkady P
authors.initials: AP
authors.lastname: Sinitsyn
mesh_terms: D000595:Amino Acid Sequence; D002236:Carbohydrate Conformation; D006026:Glycoside Hydrolases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006031:Glycosylation; D008358:Mannose / Q000737:chemistry; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010407:Penicillium / Q000201:enzymology*; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D011134:Polysaccharides / Q000737:chemistry / Q000378:metabolism; D011487:Protein Conformation; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D010455:Peptides; D011134:Polysaccharides; D006026:Glycoside Hydrolases; C019713:alpha-N-arabinofuranosidase; D008358:Mannose
keywords: Glycoproteins; High-mannose glycans; MALDI-TOF mass spectrometry; N-Glycosylation; Penicillium canescens; α-l-Arabinofuranosidase