AI Article Synopsis
- A glycosylated form of recombined human IGF-I was produced in yeast, containing only mannose as its carbohydrate component.
- Mild alkaline treatment allowed for the release and separation of all oligosaccharide chains from the glycoprotein using gel-permeation chromatography.
- The structures of these O-linked carbohydrates were analyzed, revealing specific disaccharides and tetrasaccharides, and a new pentasaccharide was identified in the yeast cell wall mannoproteins.
Article Abstract
A glycosylated form of recombinant human insulin-like growth factor I (IGF-I) expressed in Saccharomyces cerevisiae was shown to contain mannose as the only carbohydrate constituent. All oligosaccharide chains of the glycoprotein could be released by mild alkaline treatment, and separated from the protein by gel-permeation chromatography on Bio-Gel P-2. The structures of these O-linked carbohydrate chains were determined by 500-MHz 1H-NMR spectroscopy, affording the disaccharide Man alpha 1-2Man as the major component and the tetrasaccharide Man alpha 1-3Man alpha 1-2Man alpha 1-2Man as a minor component. Reference oligosaccharides were prepared from mannoproteins released from the cell wall of S. cerevisiae X2180 (alpha-wild type). In addition to previously reported structures, ranging from mannose to mannotetraose, the pentasaccharide Man alpha 1-3Man alpha 1-3Man alpha 1-2Man alpha 1-2Man was identified in the cell wall mannoprotein.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1016/0014-5793(89)80442-9 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
A novel beta-N-acetyl-D-hexosaminidase from the insect Ostrinia furnacalis (Guenée).
FEBS J
November 2008
Department of Bioscience and Biotechnology, Dalian University of Technology, China.
Exploiting specific targets is of specific interest in developing eco-friendly pesticides. We isolated, purified and characterized a novel beta-N-acetyl-D-hexosaminidase (OfHex1) from the fifth instar larva integument of the Asian corn borer, Ostrinia furnacalis (Guenée). OfHex1 was purified 1468-fold to homogeneity with an activity yield of 20% by four column chromatography steps.
View Article and Find Full Text PDFCharacterization of N- and O-linked glycosylation of recombinant human bile salt-stimulated lipase secreted by Pichia pastoris.
Glycobiology
March 2004
Wadsworth Center C-547, New York State Department of Health, PO Box 509, Albany, NY 12201-0509, USA.
Recombinant human bile salt-stimulated lipase (hBSSL) was expressed in and secreted by Pichia pastoris, an organism exploited for the large-scale production of recombinant (glyco)proteins by bioprocessing technology. The 76.3-kDa glycoprotein was associated with 75-80 Man and a small amount of GlcNAc.
View Article and Find Full Text PDFInteractions of substrate with calreticulin, an endoplasmic reticulum chaperone.
J Biol Chem
February 2003
Molecular Biophysics Unit, Indian Institute of Science, Bangalore 560012, India.
Calreticulin is a molecular chaperone found in the endoplasmic reticulum in eukaryotes, and its interaction with N-glycosylated polypeptides is mediated by the glycan Glc(1)Man(7-9)GlcNAc(2) present on the target glycoproteins. Here, we report the thermodynamic parameters of its interaction with di-, tri-, and tetrasaccharide, which are truncated versions of the glucosylated arm of Glc(1)Man(7-9)GlcNAc(2), determined by the quantitative technique of isothermal titration calorimetry. This method provides a direct estimate of the binding constants (K(b)) and changes in enthalpy of binding (Delta H(b) degrees ) as well as the stoichiometry of the reaction.
View Article and Find Full Text PDFThe role of the GlcNAc(beta)1,2Man(alpha)- moiety in mammalian development. Null mutations of the genes encoding UDP-N-acetylglucosamine:alpha-3-D-mannoside beta-1,2-N-acetylglucosaminyltransferase I and UDP-N-acetylglucosamine:alpha-D-mannoside beta-1,2-N-acetylglucosaminyltransferase I.2 cause embryonic lethality and congenital muscular dystrophy in mice and men, respectively.
Biochim Biophys Acta
December 2002
Program in Structural Biology and Biochemistry, Hospital for Sick Children, and Department of Biochemistry, University of Toronto, Ontario, Canada.
The GlcNAc(beta)1,2Man(alpha)- moiety can be synthesized by at least two mammalian glycosyltransferases, UDP-GlcNAc:alpha-3-D-mannoside beta1,2-N-acetylglucosaminyltransferase I (GnT I, EC 2.4.1.
View Article and Find Full Text PDFOccurrence of GalNAcbeta1-4GlcNAc unit in N-glycan of royal jelly glycoprotein.
Biosci Biotechnol Biochem
September 2002
Department of Bioresources Chemistry, Faculty of Agriculture, Okayama University, Japan.
Elsewhere, we characterized the structure of twelve N-glycans purified from royal jelly glycoproteins (Kimura, Y. et al., Biosci.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!