Double first! First synchrotron user from the University of Zululand solves partial structure of the Schistosomiasis (Bilharzia) G4LZI3 universal stress protein

In a ‘double first’, Dr Priscilla Masamba, has become the first University of Zululand student to use the UK’s National Synchrotron Light Source, Diamond, and solve the partial structure of a protein from Schistosoma mansoni. With access to the synchrotron made possible by GCRF START, Priscilla employed sophisticated robotic instruments and macromolecular X-ray crystallography techniques remotely from South Africa to solve the partial structure of the G4LZI3 universal stress protein – a protein regarded as a target for novel medicines for treating the disease Schistosomiasis. The experiments took place in February 2020, using the Diamond’s I04-1 beamline.

Schistosomiasis is an acute and chronic disease caused by parasitic worms (schistosomes) endemic in more than 78 countries with an estimated 4 million people infected in South Africa alone. The disease requires an intermediate host, the freshwater snail Bulinus africanus, and occurs most often in rural areas where people become infected during routine agricultural, domestic, occupational, and recreational activities which expose them to infested water.

Only one drug, Praziquantel, is available to treat Schistosomiasis leaving people vulnerable to schistosome resistance and this treatment is only partially effective in treating adults.  The aim of Priscilla ’s research is therefore to generate insights for the design of alternative treatment regimen targeting specific stages during the developmental cycle of the schistosome.

Describing the experiments at Diamond as “close to a cool sci-fi movie,” Dr Masamba was able to control the sophisticated instruments on I04-1 beamline and collect data in real time from the University of Cape Town’s (UCT’s) Aaron Klug Centre for Imaging and Analysis – established as a GCRF START Centre of Excellence for structural biology research.

“Remote data collection at Diamond was so exciting!” Dr Masamba explains, “I could literally control and see a robot that was thousands of miles away on the other side of the world, mount a microscopic crystal (sample) within the firing line of a powerful X-ray beam, and determine the amount of energies released by light emitted from the sample caused by incident X-ray beams, and all of this while working from the laboratory in Cape Town. I didn’t need to get in a plane to achieve the one of the most imperative steps in the crystallography process! The whole experience provided me with rare exposure to the world of X-ray crystallography, impacting my view of science in a spectacular way.”

Proteins are thermodynamically and kinetically responsible for all biochemical processes that occur, and are therefore responsible for coordinating, regulating and dictating numerous metabolic functions. Exposure of the Schistosome parasite to extreme conditions during its developmental stage triggers the expression of heat shock proteins and universal stress proteins, of which the G4LZI3 USP has been identified as a potential druggable target for the development of alternative treatments (schistosomicides). Techniques like X-ray crystallography can provide insight, not only into the composition of these biomolecules, but also into their various interactions with other compounds and their roles in biological mechanisms, an imperative foundation for rational drug design and development.

Before the experiments took place, diffraction of the crystals was first checked at UCT using a diffractometer. Crystals from these conditions were then flash-frozen in liquid nitrogen and shipped to the Diamond synchrotron to be used as samples.

The solved structure of the S. mansoni G4LZI3 is a success story for the University of Zululand, a small resource-constrained university in the rural part of KwaZulu-Natal Province of South Africa. The University of Zululand lacked the resources required for Dr Masamba to achieve all her objectives for her PhD, which meant the collaboration through START in order to carry out the experiments needed was imperative both professionally and personally.

Priscilla is thankful for the guidance and mentoring from her PhD supervisor, Professor Abidemi Paul Kappo, who heads up the Biotechnology and Structural Biology (BSB) Research Group in the Department of Biochemistry and Microbiology at the University of Zululand, and from START Principal Investigator, Professor Trevor Sewell, of UCT’s Aaron Klug Centre for Imaging and Analysis, both of whom helped Priscilla overcome various challenges.

“I have been able to learn and cultivate scarce, critical and sought-after skills here in Africa in the fields of bioinformatics and drug discovery, molecular biology and especially, structural biology,” says Dr Masamba. “These include gene cloning, recombinant protein expression and purification, as well as characterisation of proteins. This has not been an easy task because I am from an underrepresented group in science as a black female and study at a historically-disadvantaged and resource-constrained institution.”

An important objective of the START programme is to increase the number of structural biologists in similar less developed universities in South Africa and across the continent. This can present complex challenges, not least because many students are ill-equipped for work in the field of structural biology.

“A key concept behind the creation of the START Centre of Excellence at UCT’s Aaron Klug Centre for Imaging and Analysis, for example, is to provide the necessary infrastructure to enable senior students and staff at South Africa’s historically disadvantaged universities to access both the human and material resources necessary to overcome the difficulties and determine protein structures,” Professor Sewell says. “We count the collaboration with Professor Paul Abidemi Kappo and Dr Masamba as a major success in this respect.”

This collaboration between Prof. Kappo and Prof. Sewell was enabled by GCRF START with Prof. Sewell providing the technological resource for the G4LZI3 structural biology project, as well as the linkage to Diamond.

“Above all, Professor Sewell’s enthusiasm to train and develop a “critical mass” of students in Structural Biology is second to none,” Prof. Kappo says. “This has been a joint effort and a model of national and international collaboration. In addition to the technological resources through UCT and linkage with Diamond in the UK, funding for this project was provided by the National Research Foundation (NRF) of South Africa through a doctoral bursary awarded Dr Masamba. It is expected that structure-guided drug discovery for schistosomiasis will be the concluding part the project.”

About Dr Priscilla Masamba

Born to Congolese parents in the DR Congo, Dr Masamba lived in the UK and Zimbabwe as a child, before moving to South Africa where she matriculated and studied for her first degree in Biological Sciences at Walter Sisulu University, Mthatha. Thereafter, Priscilla joined the Biotechnology and Structural Biology (BSB) Research Group in the Department of Biochemistry and Microbiology at the University of Zululand headed by Prof Abidemi Paul Kappo and registered under his tutelage for a BSc (Hons) degree, followed by an MSc and later a PhD in Biochemistry. Priscilla’s desire is to continue in the path of macromolecular X-ray crystallography of proteins through the NRF Postdoctoral Fellowship in Structural Biology at the University of Johannesburg.

Acknowledgements

Dr Priscilla Masamba extends a special thanks to Dr Brandon Weber (UCT), Dr Phillip Venter (UCT), Kaylene Baron (UCT), and Ndibonani Qokoyi (University of Zululand) who were involved in different ways in the production, purification and crystallisation of the G4LZI3 protein, as well as in data collection.