Your browser doesn't support javascript.
loading
Glycan node profiling of soluble and membrane glycoproteins in whole cell lysates.
Aguilar Díaz de León, Jesús S; Cruz Villarreal, Jorvani; Kapuruge, Erandi P; Borges, Chad R.
Afiliação
  • Aguilar Díaz de León JS; School of Molecular Sciences and the Biodesign Institute - Center for Personalized Diagnostics, Arizona State University, P.O. Box 876401, Tempe, AZ, 85287, USA.
  • Cruz Villarreal J; School of Molecular Sciences and the Biodesign Institute - Center for Personalized Diagnostics, Arizona State University, P.O. Box 876401, Tempe, AZ, 85287, USA.
  • Kapuruge EP; School of Molecular Sciences and the Biodesign Institute - Center for Personalized Diagnostics, Arizona State University, P.O. Box 876401, Tempe, AZ, 85287, USA.
  • Borges CR; School of Molecular Sciences and the Biodesign Institute - Center for Personalized Diagnostics, Arizona State University, P.O. Box 876401, Tempe, AZ, 85287, USA. Electronic address: chad.borges@asu.edu.
Anal Biochem ; 680: 115317, 2023 11 01.
Article em En | MEDLINE | ID: mdl-37699507
ABSTRACT
Glycan node analysis (GNA) is a molecularly bottom-up glycomics technique based on the relative quantification of glycan linkage-specific monosaccharide units ("glycan nodes"). It was originally applied to blood plasma/serum, where it detected and predicted progression, reoccurrence, and survival in different types of cancer. Here, we have adapted this technology to previously inaccessible membrane glycoproteins from cultured cells. The approach is facilitated by methanol/chloroform precipitation of cell lysates and a "liquid phase permethylation" (LPP) procedure. LPP gave better signal-to-noise, yield and precision for most of the glycan nodes from membrane glycoproteins/glycolipids than the conventional solid phase permethylation approach. This GNA approach in cell lysates revealed that specific glycan features such as antennary fucosylation, N-glycan branching, and α2,6-sialylation were elevated in hepatocellular carcinoma (HepG2) cells relative to leukemia cells (THP-1 and K562) and normal donor PBMCs. Additional nodes commonly associated with glycolipids were elevated in the leukemia cells relative to HepG2 cells and PBMCs. Exposure of HepG2 cells to a fucosyltransferase inhibitor resulted in a significant reduction in the relative abundance of 3,4-substituted GlcNAc, which represents antennary fucosylation-providing further proof-of-concept that downregulation of glycosyltransferase activity is detected by shifts in glycan node expression-now detectable in membrane glycoproteins.
Assuntos
Palavras-chave

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Leucemia / Clorofórmio Limite: Humans Idioma: En Revista: Anal Biochem Ano de publicação: 2023 Tipo de documento: Article País de afiliação: Estados Unidos

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Leucemia / Clorofórmio Limite: Humans Idioma: En Revista: Anal Biochem Ano de publicação: 2023 Tipo de documento: Article País de afiliação: Estados Unidos