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Hacettepe University

3 ARTICLES PUBLISHED IN JoVE

Biochemistry

N-glycan Profiling of Glycoproteins by Hydrophilic Interaction Liquid Chromatography with Fluorescence and Mass Spectrometric Detection

Haci Mehmet Kayili¹, Bekir Salih²

¹Department of Biomedical Engineering, Karabuk University, ²Department of Chemistry, Hacettepe University

N-glycan profiling of glycoproteins is essential for discovering novel biomarkers and understanding glycan functions in cellular events. Additionally, N-glycan analysis of protein biopharmaceuticals is very important for human use. In this current article, a high-throughput strategy for identifying and quantifying N-glycan structures was presented using the HILIC-FLD-MS/MS technique.

Biochemistry

Recombinant Production of Bifidobacterial Endoglycosidases for N-glycan Release

Berfin Sucu¹, Ayşe Bayraktar^1,2, Hatice Duman¹, Ayşenur Arslan¹, Merve Kaplan¹, Melda Karyelioğlu¹, Eda Ntelitze¹, Taner Taştekin¹, Seray Yetkin¹, Melih Ertürk², Steven A. Frese^3,4, Bethany M. Henrick^4,5, Hacı Mehmet Kayili⁶, Bekir Salih⁷, Sercan Karav¹

¹Department of Molecular Biology and Genetics, Çanakkale Onsekiz Mart University, ²Uluova Dairy Farm, ³Department of Nutrition, University of Nevada, ⁴Department of Food Science and Technology, University of Nebraska, ⁵Evolve BioSystems Inc., ⁶Department of Biomedical Engineering, Karabuk University, ⁷Department of Chemistry, Hacettepe University

Bifidobacteria possess a unique genomic capability for N-glycan cleavage. Recombinantly producing these enzymes would be a promising novel tool to release bioactive N-glycans from glycoprotein-rich substrates such as colostrum.

Biochemistry

Cellular Membrane Affinity Chromatography Columns to Identify Specialized Plant Metabolites Interacting with Immobilized Tropomyosin Kinase Receptor B

Zekiye Ceren Arituluk^1,2, Bishnu Adhikari¹, Urmila Maitra¹, Caroline Goodman¹, Lukasz M. Ciesla¹

¹Department of Biological Sciences, The University of Alabama, ²Department of Pharmaceutical Botany, Faculty of Pharmacy, Hacettepe University

The protocol describes the preparation of cell membrane affinity chromatography (CMAC) columns with immobilized cell membrane fragments containing functional transmembrane tropomyosin kinase receptor B proteins. The use of CMAC columns in the identification of specialized plant metabolites interacting with these receptors and present in complex natural mixtures is also explained.