Taxonomic group: fungi / Ascomycota
(Phylum: Ascomycota)
The structure was elucidated in this paperPublication DOI: 10.1002/ejlt.201200440Journal NLM ID: 100961908Publisher: Weinheim, Germany: Wiley-VCH
Correspondence: carriere
imm.cnrs.fr
Institutions: CNRS, Aix-Marseille Université, Enzymologie Interfaciale et Physiologie de la Lipolyse, Marseille, France, University of Sfax, Laboratory of Biochemistry and Enzymatic Engineering of Lipases, Sfax, Tunisia, CNRS, Aix-Marseille Université, Architecture et Fonction des Macromolécules Biologiques, Marseille, France
The LIP2 lipase from the yeast Yarrowia lipolytica (YLLIP2) is assumed to be a good drug candidate for enzyme replacement therapy in patients with pancreatic exocrine insufficiency. Understanding and improving its biochemical properties are essential for its oral administration. YLLIP2 is a highly glycosylated protein, with glycan chains accounting for about 13% of the molecular mass of the native protein. Two potential N-glycosylation sites (N113IS and N134NT) can be identified from YLLIP2 amino acid sequence. YLLIP2 mutants with single (N113Q or N134Q) or combined (N113Q/N134Q) substitutions of these glycosylation sites were expressed in the yeast Pichia pastoris, purified and characterized. Lipase specific activity and adsorption at the lipid–water interface were found to be decreased in the absence of N-glycosylation. It was thus shown that the glycosylated enzyme had a better ability to bind and penetrate a DLPC monolayer than the non-glycosylated N113Q/N134Q mutant. Comparison of wild-type glycosylated and non-glycosylated YLLIP2 shows that the N-glycosylation clearly contributes to the high stability of YLLIP2 in the presence of pepsin in vitro, and to a lower extent in the presence of chymotrypsin. The X-ray structure of the YLLIP2 N113Q/N134Q double mutant was obtained at 2.6 Å resolution and was found to be identical to that of wild-type YLLIP2, with the lid in a closed conformation. Glycosylation is therefore not essential for a proper folding of YLLIP2.
crystal structure, enzyme, N-glycosylation, lipase, proteolysis
Structure type: monomer
Location inside paper: Fig. 6
Aglycon: (->4) Asn-X-Ser/Thr (YLLIP2)
Compound class: N-glycan
Contained glycoepitopes: IEDB_135813,IEDB_137340,IEDB_141807,IEDB_151531
Methods: amino acid analysis, MALDI-TOF, N-terminal sequence analysis
Biosynthesis and genetic data: genetic data
Comments, role: N-glycan attached to protein amino-acids with following notations: Q113, Q134
3D data: conformation data
NCBI Taxonomy refs (TaxIDs): 4922Reference(s) to other database(s): GTC:G49108TO
Show glycosyltransferases
There is only one chemically distinct structure:
Found 
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(later renamed to: Komagataella pastoris N113Q/N134Q)