A journey in parasite glycobiology and scientific infrastructure: 35 years of the Ferguson Lab in Dundee (1988–2023)

Mike is not retiring, but he is closing his research laboratory this summer. Here he reflects on what his lab has achieved, and some of the contributions he has made to the scientific infrastructure that supports WCAIR and Life Sciences in general at the University of Dundee.

The science

Recruited by Philip Cohen, Mike Ferguson arrived in Dundee from Oxford, along with Steve Homans, in 1988 Steve, a superb NMR expert, Mike and many collaborators from around the world spent several years “structure busting” – determining the primary and 3-dimensional structures of the carbohydrate components of the glycoproteins and glycolipids that dominate parasite surfaces – such as the variant surface glycoproteins (VSGs) and procyclins of T brucei, surface mucins of T cruzi and the lipophosphoglycans (LPGs) and proteophosphoglycans (PPGs) of leishmania Mike’s lab has continued this function up to the present day, developing along the way many analytic methodologies. Arguably, their heyday for structure-busting was the late 80s and 90s when they defined the surface molecular architectures of various parasite life cycle stages This work has provided a framework with which to understand parasite survival, lifestyles and tropisms in the host and vector and “images” of these parasite cell surfaces are now taken for granted.

This period was also marked by determining the structures of glycosylphosphatidylinositol (GPI) anchors from across eukaryotic evolution – defining the conserved and species/tissue/life cycle stage-specific structural features of the GPI family of molecules.

Structures are a prerequisite for studying biosynthesis, and understanding biosynthesis is a prerequisite for exploiting parasite biology for therapeutic gain Mike’s lab contributed to the detail of GPI anchor biosynthesis in African trypanosomes, leishmania and fungal pathogens and, using radiolabelling of whole cells and cell-free systems, demonstrated exploitable differences between parasite and human GPI pathways and provided genetic and chemical validation of their drug target potential

Probing the substrate and inhibitor specificities of the enzymes of the GPI pathway required access to GPI sub-structures and analogues thereof One of the challenges of working on a newly discovered pathway is that there are no reagents – let alone kits. In collaboration with Prof John Brimacombe, a superb synthetic carbohydrate chemist in our then Chemistry Department, Mike and colleagues set about synthesising unique GPI molecules and, with Andrei Nikolaev, expanded this into leishmania LPG sub-structures This era of chemical biology (much of it performed before that term was coined) was extremely productive and set this interdisciplinary team apart in the field.

Eventually, Mike’s team turned to protein N-glycosylation in trypanosomes Initial studies on the N-glycans of T brucei VSGs had suggested these were not so different to host N-glycans. However, they became interested in the “gunk” that fills the flagellar pocket of bloodstream form T brucei and found that it is made predominantly of unique giant poly-N-acetyllactosamine (PNAL) containing N-glycans With interest piqued, his team investigated N-glycan biosynthesis in T brucei and discovered that, uniquely, it has two N-glycosylation systems running in parallel – one to add complex N-glycans to glycosylation sites in acidic environments on the glycoprotein surface and another to add oligomannose structures to the remaining sites Using machine learning, they built an algorithm to predict the kinds of N-glycan present on every putative glycosylation site in the organism The unusual oligosaccharyltranferases of trypanosomes are currently being exploited in biotechnology to increase the beneficial glycosylation of recombinant glycoproteins

As with all research, one thing leads to another and one of the perplexing issues implicit in the GPI and N-glycan structures Mike’s team had determined in trypanosomes was which glycosyltransferases (GTs) were doing what?

The parasite genome is devoid of several canonical GT families and replete with members of a trypanosomatidunique one: family GT67 They therefore undertook a painstaking mapping of GT gene to GT function by constructing GT mutants and determining the biochemical lesions in GPI and/or N-glycan structures in bloodstream and procyclic form parasites. This required the development of sophisticated analytical strategies The results were worth it, and we now understand the GT67 family and its sub-families and what they do, and in the process provided the first evidence for convergent evolution in GT enzymes

One recent foray into a non-GT67 GT in T brucei (and in collaboration with Steve Beverley in L major) was to study a GT11 fucosyltransferase (FUT1) This turns out to be located in, and essential for, the parasite mitochondrion This is a radical discovery and much remains to be uncovered about mitochondrial glycosylation in general and its therapeutic potential for kinetoplastid parasites. All protein and lipid glycosylation requires nucleotide sugars as the direct or indirect donors of glycosylation reactions, making them key essential metabolites Mike’s team originally took an analytical approach by developing an LC-MS/MS method to detect and quantify nucleotide sugars (an approach later modified for mRNA cap structures) This showed GDP-fucose in all trypanosomatids, which led them to discover the trypanosomatid mitochondrial FUT1s, and stimulated them to define the biotransformations in T. brucei that make UDP-glucose, UDP-galactose, UDP-N-acetylglucosamine, GDP-mannose and GDP-fucose. Over several years, they cloned, expressed, characterised, crystallised and localised most of the thirteen enzymes involved and, crucially, discovered that all of them are localised to the glycosomes in bloodstream and procyclic form parasites This led them to discover and characterise an entirely new class of nucleotide sugar transporter, necessary to move UDP-sugars made inside the glycosomes into the cytoplasm

Throughout, Mike’s group have been heavy users of mass spectrometry, and this led Mike to set up and develop, with other colleagues like Angus Lamond and Doreen Cantrell, the School of Life Sciences proteomics facility. Open to all, it is heavily and productively used by many groups – not least Susan Wyllie’s superb mode-of-action team who have also added equipment to it. Mike’s team have contributed several fundamental proteomic data sets to the parasitology community including the first bloodstream/procyclic form proteome and phosphoproteome comparisons and comprehensive proteincomplex, cell-cycle, protein turnover and glycoproteome data.

Altogether, Mike’s lab in Dundee has published 233 peer-reviewed articles, graduated 28 PhD students and trained over 50 post-doctoral scientists.

His longest collaborator has been Lucia Güther, also his life-partner, who has played such a key role in performing original research and organising and coordinating the lab No amount of thanks are enough! Other key contributors, collaborators and fellows in the lab over the years include Angela Mehlert, Malcom McConville, Wayne Masterton, Jane Thomas-Oates, Sylvain Cottaz, Igor Almeida, Alvaro Acosta-Serrano, Terry Smith, Art Crossman, Abdel Atrih, Francoise Routier, Luis Izquierdo, Karina Marino, Mick Urbaniak, Sabine Kuettel, Manuela and Seb Damerow, Sam Duncan, Rupa Nagar and Michele Tinti. Huge thanks to all, and to Dougie Lamont and his team for managing the proteomics facility so brilliantly

Dr Lucia Guther

Scientific infrastructure and recruitment

Mike was the first molecular parasitologist to arrive in Dundee. He was able to convince Alan Fairlamb to move here in 1995 from the London School of Hygiene and Tropical Medicine, as well as David Horn from the same place and Mark Field from Cambridge in 2013 and Mattie Pawlowic from the U S in 2017 Alan’s move, as well as that that of Angus Lamond, was instrumental in Philip Cohen convincing the Wellcome Trust to invest £10 million into the Wellcome Trust Building which opened in 1997. Alan also attracted Bill Hunter to bring X-ray crystallography to Dundee from Manchester, and Alan, Mike and Bill shared a vision for advancing drug discovery for neglected tropical diseases

The next phase required more building – Mike and Philip Cohen jointly led the campaign to fund and build the Sir James Black Centre, which opened in 2006 This included the first Drug Discovery Unit (DDU) laboratories and Mike and Alan initiated drug discovery the same year with an extraordinary strategic award from Wellcome, and by making key initial appointments of Julie Frearson and Ian Gilbert and then subsequently Paul Wyatt. Kevin Read was recruited shortly thereafter and David Gray succeeded Julie Frearson in 2010 The DDU team has been enormously successful over the years and is now 130-strong with a turnover in excess of £12m pa, five drugs in clinical trials and nine assets licenced to BioPharma companies The translational capabilities of the DDU were also central to the establishment of the Wellcome Centre for Anti-Infectives Research in 2017, one of only fifteen Wellcome Centres

The burgeoning success of the DDU meant that it was outgrowing its laboratories and, in 2009, Mike initiated the fund raising for, and the design of, the Discovery Centre for Translational and Interdisciplinary Research. This £25m development opened in 2014 and allowed the DDU to double in size It also provided purpose-built accommodation for our proteomics facility (one of the largest in the world).

Since 2017, Mike and Morag Martin have been involved with the Tay Cities Deal (TCD), shaping and justifying the £25m TCD investment in Growing the Tay Cities BioMedical Cluster, raising additional funds from the Garfield Western and Wolfson Foundations and making the case for University of Dundee strategic investment. Construction of the Life Sciences Innovation Hub began on the Dundee Technopole in March 2023 and will complete in 2024 It will provide laboratory and office space for spin-out companies, reversing the current trend for the University’s highly-invested companies to move elsewhere – thus retaining jobs and opportunities in this economically depressed region Together with Alessio Ciulli’s Centre for Targeted Protein Degradation, this is the first step towards developing a wider Life Sciences Innovation District for the prosperity of the region

Summing up

In summarising his career so far, Mike said: “It has been a privilege to work with so many talented scientists and to collectively add something to the canon of molecular parasitology and eukaryotic biology. It has also been a pleasure, if somewhat time consuming, to help provide facilities and infrastructure for the next generation of scientists to thrive. Dundee has been a fantastic home and work environment for me, and I hope it is and will be for many others for many years to come.”