Preface

Glycoprotein Analysis is a comprehensive overview of contemporary methods used for studying the structures, dynamics, interactions and assembly of glycoproteins. The overarching aim of this book is to provide an all-inclusive view of glycoprotein analysis, incorporating orthogonal and complementary techniques in a single volume, as opposed to focusing on a single methodological approach as done previously in subdisciplines of glycobiology. This volume is part of the New Developments in Mass Spectrometry series, and although mass spectrometry (MS) is fundamentally important in glycobiology, it is limited in its ability to characterise glycoproteins. Therefore, we have included orthogonal methods, such as NMR, molecular dynamics and lectin array technologies. Each chapter focuses on a specific advanced technique for examining glycoproteins – from methods proficient in characterising structures to those capable of understanding their functional interactions. The approaches covered herein vary, with many recently emerging and proving groundbreaking for glycoprotein analysis. The topics and applications are highly relevant across academia and industry, ranging from basic research to advanced biophysical experiments. Our goal is for this book to be a valuable resource for a diverse audience for years to come.

Often, we consider glycomics to trail similar -OMICs fields, a viewpoint arguably based on challenges associated with structural analyses. By comparison, third-generation DNA sequencing methods have achieved single-molecule detection (e.g. nanopore sequencing), and single-cell proteomes are routinely measured using commercial mass spectrometers. Optimistically, glycoscience will follow suit. However, a principal challenge posed by glycoproteins rests in their intrinsic structural heterogeneity derived from non-templated glycoconjugate biosynthesis. This obstacle has been referred to as the “glycosylation problem” in structural biology. Despite significant advances in cryogenic electron microscopy, glycoproteins remain effectively intractable, as glycans are seldom resolved beyond the initial few conserved residues attached to the peptide backbone. Therefore, in the context of the cryo-EM “resolution revolution” nearly ten years ago, and its subsequent widespread growth, glycoproteins are arguably excluded. Nonetheless, tools for studying protein glycosylation have advanced with the emergence of innovative methods capable of bridging the gaps between genomics, proteomics, transcriptomics, metabolomics and glycomics.

From a technical perspective, the need for new methods has been highlighted in several reports, including CarboMet’s Glyco 2030: A Roadmap for Glycoscience in Europe (2019) and National Research Council’s Transforming Glycoscience: A Roadmap for the Future (2012). Likewise, the need for analytical methods is explicitly stated in the Essentials of Glycobiology textbook (Chapter 60, Future Directions in Glycoscience). Logically, although technical advances are important, it is equally imperative to make new methodologies accessible to non-experts. This will not only ensure growth in glycoscience but also encourage a greater appreciation of glycans in biology and medicine. Undoubtedly, the most critical consideration looking forward is educating emerging researchers who will lead the field in the future. This sentiment was echoed in the 2016 perspective “Training the next generation of biomedical investigators in glycosciences”, which stressed the lack of glycobiology in higher education and the need to establish training centres to integrate glycobiology into mainstream life science curricula.

Glycobiology has experienced periods of accelerated growth. For example, in the 1980s, advances in oligosaccharide separation and detection, as well as the availability of carbohydrate standards and enzymes, made sequencing glycans possible. It was in 1988 when Raymond Dwek coined the term “glycobiology”, and influenced the Oxford English Dictionary to adopt the term in their 1992 edition, helping to mark the field as a distinct discipline. One could argue that glycobiology is experiencing a period of development at present, driven not only by exciting new technologies but also by the recognised role of glycans in clinical medicine. The COVID pandemic brought glycans into the spotlight, with images of the spike glycoprotein often (but not always) shown with its glycan shield. Glycobiology is an integral part of vaccine safety and efficacy. Accordingly, this book begins with a chapter covering SARS-CoV-2 glycosylation and how it relates to glycan structural analysis more generally.

Our understanding of glycosylation in health and disease is consistently progressing, from identifying new functions across biological processes in cellular signalling and immunity to pathology and infection. Today, glycobiology continues to be a dynamic and integral area of research, intersecting with various disciplines and contributing to significant biomedical advancements. Thus, the analysis of glycoproteins is a cornerstone in biochemistry and molecular biology, with significant implications in diagnostics, biotherapeutics and drug development. Confidently, we can view the challenges in glycoscience as opportunities, many of which are being addressed through technical advances that are covered in this book by world-leading experts.

Weston B. Struwe

Department of Biochemistry

Kavli Institute for NanoScience Discovery

University of Oxford, OX1 3QU, UK