title: Synthetic Glycoforms Reveal Carbohydrate-Dependent Bioactivity of Human Saposin D.
abstract: The main glycoforms of the hydrophobic lysosomal glycoprotein saposin D (SapD) were synthesized by native chemical ligation. An approach for the challenging solid-phase synthesis of the fragments was developed. Three SapD glycoforms were obtained following a general and robust refolding and purification protocol. A crystal structure of one glycoform confirmed its native structure and disulfide pattern. Functional assays revealed that the lipid-binding properties of three SapD glycoforms are highly affected by the single sugar moiety of SapD showing a dependency of the size and the type of N-glycan.
authors.affiliation: Bioorg. Chemie, Gebäude NWI, Universität Bayreuth, 95440, Bayreuth, Germany.
authors.forename: Christopher G F
authors.initials: CGF
authors.lastname: Graf
authors.affiliation: Bioorg. Chemie, Gebäude NWI, Universität Bayreuth, 95440, Bayreuth, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Schulz
authors.affiliation: Bioorg. Chemie, Gebäude NWI, Universität Bayreuth, 95440, Bayreuth, Germany.
authors.forename: Marina
authors.initials: M
authors.lastname: Schmälzlein
authors.affiliation: Bioorg. Chemie, Gebäude NWI, Universität Bayreuth, 95440, Bayreuth, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Heinlein
authors.affiliation: Bioorg. Chemie, Gebäude NWI, Universität Bayreuth, 95440, Bayreuth, Germany.
authors.forename: Manuel
authors.initials: M
authors.lastname: Mönnich
authors.affiliation: Bioorg. Chemie, Gebäude NWI, Universität Bayreuth, 95440, Bayreuth, Germany.
authors.forename: Lukas
authors.initials: L
authors.lastname: Perkams
authors.affiliation: Bioorg. Chemie, Gebäude NWI, Universität Bayreuth, 95440, Bayreuth, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Püttner
authors.affiliation: Bioorg. Chemie, Gebäude NWI, Universität Bayreuth, 95440, Bayreuth, Germany.
authors.forename: Irene
authors.initials: I
authors.lastname: Boos
authors.affiliation: Bioorg. Chemie, Gebäude NWI, Universität Bayreuth, 95440, Bayreuth, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Hessefort
authors.affiliation: Bioorg. Chemie, Gebäude NWI, Universität Bayreuth, 95440, Bayreuth, Germany.
authors.forename: Jose Nelson
authors.initials: JN
authors.lastname: Lombana Sanchez
authors.affiliation: Department of Biochemistry, Universität Bayreuth, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Weyand
authors.affiliation: Department of Biochemistry, Universität Bayreuth, Germany.
authors.forename: Clemens
authors.initials: C
authors.lastname: Steegborn
authors.affiliation: LIMES Institute, Universität Bonn, Germany.
authors.forename: Bernadette
authors.identifier: 0000-0002-4838-4109
authors.initials: B
authors.lastname: Breiden
authors.affiliation: LIMES Institute, Universität Bonn, Germany.
authors.forename: Kerstin
authors.identifier: 0000-0003-3152-2269
authors.initials: K
authors.lastname: Ross
authors.affiliation: LIMES Institute, Universität Bonn, Germany.
authors.forename: Günter
authors.identifier: 0000-0002-0203-8331
authors.initials: G
authors.lastname: Schwarzmann
authors.affiliation: LIMES Institute, Universität Bonn, Germany.
authors.forename: Konrad
authors.identifier: 0000-0003-2452-8654
authors.initials: K
authors.lastname: Sandhoff
authors.affiliation: Bioorg. Chemie, Gebäude NWI, Universität Bayreuth, 95440, Bayreuth, Germany.
authors.forename: Carlo
authors.initials: C
authors.lastname: Unverzagt
mesh_terms: D002236:Carbohydrate Conformation; D002241:Carbohydrates / Q000737:chemistry*; D006031:Glycosylation; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D010316:Particle Size; D049231:Saposins / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism*
chemical_list: D002241:Carbohydrates; D049231:Saposins
keywords: glycopeptide; glycoprotein; ligation; lipid metabolism; oligosaccharide