In 2021, LP3 continued to deliver projects to its users at the maximum of its capacities and as much as allowed by the Covid-19 pandemic. LP3 continued to offer protein crystal screening at BioMAX, the X-ray macromolecular crystallography beamline of MAX IV Laboratory, to its users. This enables non-experts in protein crystallography to use X-ray crystallography at MAX IV.

In 2021, LP3 staff was involved in both undergraduate and graduate teaching, as well as national and international conferences and networks of interest, but these activities were severely limited by the Covid-19 situation.

However, as should be clear from the pages of this annual report 2021, LP3 continues to deliver value-adding services to Lund University researchers.

Also, during 2021, an external evaluation of LP3, based on the performance during the last 5 years, was done by Prof. Per Gardeström from Umeå University. In addition, the Swedish Research Council has granted financing for a five-year period (2022 – 26) for a new nationally distributed research infrastructure – Protein Production Sweden (PPS). LP3 will be running one of the PPS nodes.

Introduction

Lund Protein Production Platform (LP3) is a focal point for expertise and equipment for the entire process chain of production, purification, characterization, crystallization of proteins and their structure determination and refinement, or each individual step in the chain. LP3 is a service centre that offers customer-adapted protein production, including stable isotope-labelled proteins, crystallization of proteins and structure determination of proteins primarily for Lund University (LU), but also for the surrounding community.

Since 2018, LP3 is part of a Block Allocation Group (BAG) proposal of Lund researchers and has beamtime at the BioMAX beamline at MAX IV laboratory. LP3 does therefore also handle regular screening of user crystals at BioMAX and subsequent structure determination. LP3 is also a knowledge centre for dissemination and exchange of new technologies and ideas within protein production and protein crystallization.

The mission of LP3 is to:

• offer open service and support, primarily to researchers at Lund University, with protein production, characterization and crystallization for their research projects.
• be responsible for a common and open infrastructure for protein production and crystallization, as well as to contribute actively to the interaction of Lund University with MAX IV, European Spallation Source ERIC (ESS) and other relevant major research facilities, networks and initiatives.
• if needed, to act as a node of Lund University in a national infrastructure in the protein science area.
• develop competence and methods in the area of protein sciences.
• serve the surrounding community (closely located large infrastructures, small biotech companies, and such).
• finance part of its operations (material and machine maintenance costs) by charging user fees and to increase this part of the funding over time.

5-year evaluation

During 2021, an external evaluation of LP3 commissioned by the Dean of the Faculty of Science, based on the LP3 performance during the last five years, was done by Prof. Per Gardeström from Umeå University, with particular focus on LP3s mission goals. This point was summarized in the report as: “LP3 has fulfilled its mission in an excellent way, according to the goals formulated by Lund University. However, IT problems at the crystallization unit have to some extent been negative for the service that could be provided”.

Organisation

The infrastructure was in 2021 run by a manager (50 % full-time equivalent, senior lecturer) and in average five research engineers of whom three were involved in protein production and two primarily in protein crystallization, protein structure determination and biophysical protein characterization. In 2021, the staffing also included two experts (10 %) in microbial protein production and crystallization as well as Dr. Z. Fisher (Head of DEMAX (ESS), associate lecturer at Lund University). Four visitors were associated with LP3 in 2021, not counting the staffing of the FragMAX project.

During 2021, two research engineers left LP3 and only one position could be partially replaced, due to suddenly cancelled support to LP3 by the Exodiab research area at the Faculty of Medicine. This together with the restrictions for working due to the Covid-19 pandemic impacted of course on the deliveries and speed at which projects could be executed.

LP3 is fully equipped for protein production in E. coli and insect cells (Baculovirus Expression Vector System, BEVS). This includes flow hoods for sterile handling of cells, temperature-controlled shakers for culturing of cells (including access to temperature controlled rooms), centrifuges, cell homogenization equipment (e.g., French Press and sonicators). For purification there are several chromatography systems, including one Äkta Avant, one Äkta Pure and one Äkta Purifier System. Equipment for SDS-PAGE, Western blotting and other standard equipment for protein characterization and enzymatic activity assays is available at the centre or within close proximity. All documentation is captured using electronic lab notebooks. For crystallisation, the facility is equipped with state-of-the-art nanolitre pipetting equipment with the capability to handle lipidic cubic phases for membrane proteins, as well as a “plate hotel” with the capacity to store and automatically image plates. Tecan and a TTP (Dragonfly) liquid handling systems for the preparation of crystallization screens are also available.

For more specifics on the capabilities and services of LP3, please see “Services below”.

Placement of the infrastructure

LP3 is placed at the Department of Biology (Biology Building A, Sölvegatan 35, 22362 Lund), within the Faculty of Science at Lund University. LP3 is a separate entity within the existing administrative structure of the Department of Biology and follows the working and delegation principles of the Faculty of Science.

Leadership of the infrastructure

LP3 is governed by a board of one chairman (Prof. Anders Tunlid) and 6 members (Prof. Susanna Horsefield, Dr. Kajsa Paulsson, Prof. Mikael Akke, Dr. Kajsa Sigfridsson Clauss, Dr. Sindra Petersson Årsköld, Ida Lunga), one each from Faculty of Science (FoS), Faculty of Medicine (FoM), LTH, MAX IV laboratory and ESS (external member) and one student. The chairman is the dean or pro dean of the Faculty of Science. The daily business of the center is led by a manager (50 % full time equivalent) (Dr. Wolfgang Knecht). The manager is supported in his function by additional experts (10 % full time equivalent) (Currently Dr. Claes von Wachenfeldt (microbiological protein production and deputy manager LP3) and Prof. Derek Logan (crystallisation).

Deuteration and Macromolecular Crystallization (DEMAX)

The DEMAX platform of the European Spallation Source ERIC (ESS) is co-localized with LP3 since 2016. DEMAX and LP3 are collaborating to coordinate their efforts to develop cost-effective production of deuterated biomaterials (lipids and proteins) for neutron-based methods such as protein crystallography, neutron reflectometry, and small angle neutron scattering. The underlying agreement for this was renewed for the period 2021-2025.

FragMAX (BioMAX Fragment Screening platform)

The FragMAX platform of MAX IV (external website) is a Swedish Research Council financed project, with LP3, AstraZeneca AB and Saromics Biostructures AB as co-applicants. The project aims at setting up high throughput X-ray fragment screening (XFS) at the BioMAX beamline with LP3 as the partner for crystallization and sample preparation. This part of FragMAX is localized in LP3 laboratories since 2019 as illustrated in Figure 1.

In summary, the laboratories of LP3 are already today a physical meeting point of three organizations: LP3, MAX IV laboratory and ESS. This is in line with LP3s amibition to be seen as Gateway environment.

SWEbeams (an initiative done during 2017-2018, commissioned by Vinnova and the Swedish Research Council with the aim to create world-leading platforms for research, innovation and collaboration that enable the use of the unique potential of ESS and MAX IV.) defines, in its final report, the concept of “Instegsmiljö” – Gateway environment, which in the broadest sense describes platforms and constellations that allow preparation of reagents and/or provide expertise for the subsequent performance of experiments at synchrotrons and neutron sources, in particular of Swedish interest MAX IV laboratory and in the future ESS. Consequently, LP3 was recently mentioned as perceived by its users as a well-functioning example for a Gateway environment in the Vinnova rapport VR 2020:04 “Nationell Science Park i anslutning till forskningsanläggningarna ESS och MAX IV”

Long term strategy

A new nationally distributed research infrastructure – Protein Production Sweden (PPS) – will start up in the beginning of 2022. The Swedish Research Council has granted financing for a five-year period, until 2026. Five universities (the University of Gothenburg (host), Lund University, Karolinska Institutet, Royal Institute of Technology, and Umeå University) form the infrastructure and offer expert competence in various techniques of protein production. Researchers across Sweden will get access via a joint hub and will have the possibility to get support based on their research needs.

As the Lund node (Figure 2) LP3 will provide 3 of the 8 modules of PPS. In the context of conventional protein production modules, LP3 will provide protein expression in insect cells using the baculovirus expression vector system (BEVS). In addition, LP3 will be responsible for the production of (per)deuterated proteins for neutron scattering applications in one module (4.1) and reagents to aid in structural biology at synchrotrons in another one (4.2). With the two later modules, LP3 connects seamless to its capabilities for protein crystallisation, structure determination and X-ray aided fragment screening (XFS) at FragMAX. LP3 is thereby responsible within PPS for the goal to actively create small and specific Gateway environments for Max IV and ESS.

The PPS platform matches LP3s mission statement “if needed, to act as LU’s node in a national infrastructure in the protein science area” but also is in line with the previously formulated longterm strategy for LP3. Here we stated that in the 3 to 5 years perspective, LP3 currently works towards to establish itself as provider of:

Preferred supplier of (per)deuterated biomolecules for neutron scattering experiments

Protein Structure Determination for non-experts (non-crystallographers)

LP3 is to act as a gateway environment, which in the broadest sense describes platforms and constellations that allow preparation of reagents and/or provide expertise for the subsequent performance of experiments at synchrotrons and neutron sources, in particular of Swedish interest, MAX IV laboratory and in the ESS. These ambitions of LP3 feature two specializations that cater exclusively to use in neutron sources and/or synchrotrons.

LP3 is co-localized with and closely collaborates with the DEMAX platform of the ESS on production of deuterium labeled biomolecules for neutron scattering and the development of methods for crystal growth for neutron crystallography. Together with DEMAX, LP3 is to our knowledge the only platform in Europe that provides both labeled and unlabeled protein production services, and a pipeline directly to crystallization. The D-lab at the Institut Laue-Langevin (ILL) in France offers only support for deuterated protein production. LP3 is therefore in a unique position to assist users that want to execute biological neutron scattering experiments at existing large research infrastructures like for example the ILL in France, the ISIS Neutron and Muon Source of the STFC in the UK or in the future, the ESS in Sweden. This forms the basis of module 4.1 in PPS.

Also, LP3 capabilities to provide service in principle from “gene to structure”, in particular for non-experts in structural biology with X-ray crystallography at MAX IV are shown later in the report with a large number of projects combining protein production and structure determination for LP3 users. This forms the basis of module 4.2 in PPS, for specific production of molecular chaperones helping in co-crystallisation.

Preferred supplier of protein expression in insect cells with the BEVS in Sweden

Insect cells are particularly successful in production of large proteins and protein complexes, and can handle many post-translational modifications. In contrast to other eukaryotic expression systems, such as mammalian cells and yeast cells, they are very good for production of mammalian cytosolic proteins. For example, insect cells are widely used for expression of protein kinases. Other important factors are the ease of culturing and infecting insect cells (cost effective medium and no need for CO2 supply, in contrast to mammalian cells), and the high recombinant protein yields obtainable. A majority of proteins produced in eukaryotic expression systems and used for protein structure determination (deposited in PDB), have been produced in insect cells (Publication: Assenberg, R., et al., Advances in recombinant protein expression for use in pharmaceutical research. Curr Opin Struct Biol, 2013. 23(3): p. 393-402).

LP3 is using the BEVS to provide recombinant proteins to its users since 2013 and protein production in insect cells makes up for about 30 – 40 % of its protein production. LP3 also collaborates both in the development, as well as in implementing “best practice” for the BEVS with other protein platforms internationally. This collaboration has resulted in a publication in the Journal of Structural Biology.

LP3 is the only university platform in Sweden offering protein expression in insect cells using the BEVS. LP3's ambition in this area is to continuously improve its service to users by implementing newer and or more efficient developments of the system at LP3 within the PPS (module 3.3).

XFS together with BioMAX after the FragMAX project

LP3 ambition is to be able to offer also beyond 2022, in collaboration with the MAX IV laboratory, crystallization and sample preparation services for the FragMAX platform and contribute to its operational phase and constant development.

CryoEM

The external evaluation by Prof. Per Gardeström suggested location of CryoEM competence at LP3. This and in particular the engagement of LP3 in PPS, as also concluded in Pers report, requires a certain increase in staff compared to what is currently working at LP3 to further being able to fulfill LP3s mission goals.

In an even longer time-frame, LP3 board supported the idea of LP3 becoming a part of a future Integrative Structural Biology Center (ISBC) at Science Village (SV). The vision is to establish a Research Center in close vicinity to MAX IV and ESS in order to generate unique synergistic opportunities and stimulate strong scientific collaborations within the area of integrative structural biology.

Covid-19

In 2021 the Covid-19 situation has of course also impacted on LP3 with respect to higher absence of staff from experimental work at the laboratories due to the restrictions for work given by the Department of Biology, in particular during the first part of 2021. This also impacted on LP3 possibilities to host visitors and users to access equipment made available by LP3. The loss of productivity is estimated in the range of one to two full-time employee years. Also repair and maintenance of some equipment was severely suffering due to travel restrictions for the technical personal that would have to come from abroad/oversees, contributing to the above-mentioned loss of capacity. Furthermore, delivery shortages – long delivery times (in particular with regard to plastic materials) became obvious in 2021, forcing LP3 to build up stocks on site rather than ordering in time. While not directly impacting productivity so far, it impacted with respect to relatively higher running costs in 2021 than expected.

Due to the Covid-19 pandemic, LP3 already in 2020 prioritized work on SARS-CoV-2 related projects by for example supporting the Covid-19 action program of the ESS, resulting in the first structure determination of the non-structural-protein 10 (nsp10) from SARS-CoV-2, which was published in the International Journal of Molecular Sciences. A continuation of this work in a collaboration between LP3, DEMAX, FragMAX and University College London has delivered unique insights into the druggability of nsp10.

Additionally, LP3 became a partner in a European University Alliance for GLObal Health (EUGLOH) financed project: "Disentangling and visualising the key epitopes for neutralising antibodies against SARS-CoV-2 at the atomic level - Implications for development of vaccines against COVID-19" under the lead of Joakim Esbjörnsson (Faculty of Medicine) and participated in an application within the thematic collaborative initiative call of Lund University addressing pandemic spread with the title PandA (Pandemics and Alertness), which in 2022 will be incorporated as a stand alone theme into the new Lund University Virus Center.

Services

LP3 offers services for the entire process chain of production, purification, characterization, protein crystallization and protein structure determination and structure refinement or each individual step in the chain. LP3 can help with:

• Plasmids for protein production
• Recombinant protein production in bacterial (E. coli) or eukaryotic (insect) cells.
• Protein labeling (seleno-methionine incorporation, labeling with stable isotopes (²H, ¹³C, ¹⁵N), biotinylation, phosphorylation)
• Protein purification
• High-throughput & nanovolume protein crystallization
• Biophysical protein characterization by Size Exclusion Chromatography (SEC), Dynamic Light Scattering (DLS) and Differential Scanning Fluorimetry (DSF)
• Automated crystallization plate storage and imaging
• Protein structure determination and refinement:
  • Application for beamtime. LP3 is part of a Block Allocation Group (BAG) for BioMAX.
  • MX data collection at synchrotron beamline (BioMAX MAX IV)
  • Process x-ray data and determine and refine the structures
  • Cryo-EM screening, LP3 is part of a Block Allocation Group (BAG) for SciLifeLab Cryo-EM in Stockholm and Umeå

For details of current services and updates, please see our webpage crystallisation.

Users and projects

An overall user statistics will be reported here and a more detailed breakdown into protein production and the crystallization part will also be presented. The data will be also presented in the context of the timeframe 2016 – 2021.

Overall

53 groups used LP3 in 2021. Of these, 44 principal investigators came from Lund University and 9 were external. The distribution into different faculties and external users is presented in Figure 3. The development of user groups at LP3 since 2016 is shown in Figure 4. The principal investigators in the external user group come from the ESS, other Swedish universities (for example in 2021: Uppsala University, Umeå University, the Royal Institute of Technology, Martin-Luther University (Germany)) and industry/biotech (in 2021: two companies).

Figure 5 presents the total distribution of 118 unique users at LP3 in the timeframe of 2016 - 2021.

15 (one from the Faculty of Engineering, six form the Faculty of medicine and five from the Faculty of Science) of these user groups had at least one project at LP3 every year in this period and additional two principal investigators from the faculty of Science had at least one project in 5 out of the 6 years. This shows that LP3 has built up a basis of regular users that trust in and depend on LP3 for their supply with recombinant proteins and access to protein characterization and structure determination.

121 unique deliveries were made in 55 protein production projects and 258 protein crystallization plates were processed. Four visitors worked at LP3 for periods between a few days to up to one year or are still associated with LP3.

Seven projects are set up for both protein production and crystallization at and by LP3 with the aim of structure determination by X-ray crystallography for non-expert users. In three additional projects, LP3 does protein production with the purpose of crystallization elsewhere by the user. In another four projects, LP3 receives protein for crystallization and structure determination services by LP3. Of the total 55 projects in protein production at LP3, 10 or 18 % have a direct link to X-ray crystallography and this shows clearly a demand for LP3 capabilities to provide service in principle from “gene to structure”, in particular for non-experts in structural biology.

Protein Production

Figure 6 below shows the number and distribution of users in 2021. In Figure 7 the development of user groups is illustrated for 2016 – 2021 and in Figure 8, it is shown how many users had their first project at LP3 in protein production on a yearly basis.

For seven of the 40 principal investigators, it was the first time that they used LP3 to run a protein production project in 2021.

In total, LP3 worked with 55 protein production projects in 2021. The distribution of projects is illustrated in Figure 9. The development of the number and distribution of protein production projects is shown on a yearly basis in Figure 10.

In 2021, 121 unique deliveries were made in these 55 protein production projects and their distribution is shown in Figure 11, as well as the development of unique deliveries (for example protein batches) on a yearly basis for 2016 – 2021 is illustrated in Figure 12.

The above figures show that within protein production in 2021, 58 – 73 % of LP3s user groups, projects and deliveries are from and go to the Faculty of Science and the Faculty of Medicine. This is in the same orders as in 2020, with a range of 61 – 68 %

31 % and 16 % of all projects are connected to two areas of specialization of LP3, either the BEVS or stable isotope labelling, respectively.

Protein Crystallisation and biophysical protein characterization

A total of 258 screening plates were processed in 2021. This area of LP3 had 27 user groups. LP3 had 12 projects with the aim of structure X-ray crystallography managed by LP3 for non-expert users.

The distribution of user groups in crystallization and the development of the number and distribution of user groups since 2016 are shown in Figure 13 and 14. Traditionally and in contrast to protein production, here a higher percentage of users which come from the Faculty of Science than the Faculty of Medicine. This has however changed in 2020 (Figure 14), with now also users from the Faculty of Medicine making up the biggest user group. This is probably also triggered by an increase of users accessing equipment made available by LP3 to all LU researchers within biophysical characterization (e.g. nanoDSF, DSL) at LP3.

Crystal screening at the BioMAX Beamline

202 crystals from 13 projects in crystallography completely managed by LP3 for non-expert users or owned by users were tested by LP3 staff at the BioMAX at 11 occasions and 63 datasets were collected in total. Seven of these projects were managed by LP3 and resulted in 136 crystals and 55 datasets. Five other LP3 lead projects did not come to the stage yielding crystals, yet.

Additionally, LP3 helped expert users in crystallography in 6 projects to test 66 crystals at the BioMAX beamline resulting in 8 datasets.

FragMAX project (BioMAX Fragment Screening platform)

In 2021, due to Covid-19 restrictions and staff turnover in the FragMAX project: Tobias Krojer starting as new project leader in January and Sandesh Kanchugal as postdoc in May, activities were limited to internal project activities (including the SARS-CoV-2 nsp10 project mentioned before) and one user group visiting in autumn for crystal setup, soaking and screening.

Visibility, access, outreach

LP3 presents its services, capabilities and new developments through this website and Lund University's research portal's infrastructure pages, as well as at meetings (see below).

LP3 participates in relevant national and international networks and societies, (like Protein Production Network Sweden (PPNS), Protein Production and Purification Partnership in Europe (P4EU) and Core Technologies for Life Sciences (CTLS). Since 2019, LP3 is one of the Lund University infrastructures affiliated with the European infrastructure for translational medicine (EATRIS ERIC) in their small molecule platform. LP3 is also a member of the European Deuteration Network DEUNET.

The above engagements are for dissemination of LP3’s work as well as for the exchange and adoption of new ideas and methods into LP3. LP3 staff normally participates in seminar series, research schools and conferences. However due to the Covid-19 pandemic in 2021, LP3 staff only participated in hybrid/virtual events. These cover the virtual P4EU, DEUNET and CTLS meetings, as well as the LINXS membrane protein workshops. Oral presentations of LP3 were given at the LUBIRC Breakfast Seminar, MAX IV user days, the MAX IV structural biology workshop and the Protein Science Day in Lund. A poster presentation of LP3 was presented at the 24th Swedish Conference on Macromolecular Structure and Function.

As nearly every year, LP3 was contributing to undergraduate education as well as the Ph.D course “Protein Factories”.

Results and /or proteins produced at the facility were used in the following 2021 publications – including accepted in 2021 (in the total time 2016 – 2021, the total number of publications has thereby reached 84):

1. Aggarwal, S.; Wachenfeldt, C.V.; Fisher, S.Z.; Oksanen, E. A protocol for production of perdeuterated OmpF porin for neutron crystallography. Protein Expr Purif 2021, 188, 105954, doi:10.1016/j.pep.2021.105954.
2. Bahnan, W.; Happonen, L.; Khakzad, H.; Ahnlide, V.K.; de Neergaard, T.; Wrighton, S.; Bratanis, E.; Tang, D.; Hellmark, T.; Björck, L.; et al. Protection induced by a human monoclonal antibody recognizing two different epitopes in a conserved region of streptococcal M proteins. bioRxiv 2021, 2021.2003.2001.433494, doi:10.1101/2021.03.01.433494.
3. Bergwik, J.; Kristiansson, A.; Larsson, J.; Ekström, S.; Åkerström, B.; Allhorn, M. Binding of the human antioxidation protein α(1)-microglobulin (A1M) to heparin and heparan sulfate. Mapping of binding site, molecular and functional characterization, and co-localization in vivo and in vitro. Redox Biol 2021, 41, 101892, doi:10.1016/j.redox.2021.101892.
4. Ding, B.-J.; Wang, H.-L.; Al-Saleh, M.A.; Löfstedt, C.; Antony, B. Bioproduction of (Z,E)-9,12-tetradecadienyl acetate (ZETA), the major pheromone component of Plodia, Ephestia, and Spodoptera species in yeast. Pest Management Science 2021, n/a, doi.org/10.1002/ps.6716.
5. Ding, B.-J.; Xia, Y.-H.; Wang, H.-L.; Andersson, F.; Hedenström, E.; Gross, J.; Löfstedt, C. Biosynthesis of the Sex Pheromone Component (E,Z)-7,9-Dodecadienyl Acetate in the European Grapevine Moth, Lobesia botrana, Involving ∆11 Desaturation and an Elusive ∆7 Desaturase. Journal of Chemical Ecology 2021, 47, 248-264, doi:10.1007/s10886-021-01252-3.
6. Kristiansson, A.; Bergwik, J.; Alattar, A.G.; Flygare, J.; Gram, M.; Hansson, S.R.; Olsson, M.L.; Storry, J.R.; Allhorn, M.; Åkerström, B. Human radical scavenger α(1)-microglobulin protects against hemolysis in vitro and α(1)-microglobulin knockout mice exhibit a macrocytic anemia phenotype. Free Radic Biol Med 2021, 162, 149-159, doi:10.1016/j.freeradbiomed.2020.02.018.
7. Lassance, J.-M.; Ding, B.-J.; Löfstedt, C. Evolution of the codling moth pheromone via an ancient gene duplication. BMC Biology 2021, 19, 83, doi:10.1186/s12915-021-01001-8.
8. Nouri, M.; Latorre-Margalef, N.; Czopek, A.; Råberg, L. Cross-reactivity of antibody responses to Borrelia afzelii OspC: Asymmetry and host heterogeneity. Infect Genet Evol 2021, 91, 104793, doi:10.1016/j.meegid.2021.104793.
9. Passot, F.M.; Cantlay, S.; Flärdh, K. Protein phosphatase SppA regulates apical growth and dephosphorylates cell polarity determinant DivIVA in Streptomyces coelicolor. Molecular microbiology 2021, n/a, doi.org/10.1111/mmi.14856.
10. Petruk, G.; Puthia, M.; Petrlova, J.; Samsudin, F.; Strömdahl, A.C.; Cerps, S.; Uller, L.; Kjellström, S.; Bond, P.J.; Schmidtchen, A.A. SARS-CoV-2 spike protein binds to bacterial lipopolysaccharide and boosts proinflammatory activity. J Mol Cell Biol 2021, 12, 916-932, doi:10.1093/jmcb/mjaa067.
11. Roberts, R.E.; Yuvaraj, J.K.; Andersson, M.N. Codon Optimization of Insect Odorant Receptor Genes May Increase Their Stable Expression for Functional Characterization in HEK293 Cells. Frontiers in Cellular Neuroscience 2021, 15, doi:10.3389/fncel.2021.744401.
12. Stenström, O.; Diehl, C.; Modig, K.; Nilsson, U.J.; Akke, M. Mapping the energy landscape of protein–ligand binding via linear free energy relationships determined by protein NMR relaxation dispersion. RSC Chemical Biology 2021, 2, 259-265, doi:10.1039/D0CB00229A.
13. Venskutonytė, R.; Koh, A.; Stenström, O.; Khan, M.T.; Lundqvist, A.; Akke, M.; Bäckhed, F.; Lindkvist-Petersson, K. Structural characterization of the microbial enzyme urocanate reductase mediating imidazole propionate production. Nature communications 2021, 12, 1347, doi:10.1038/s41467-021-21548-y.
14. Wallerstein, J.; Ekberg, V.; Ignjatović, M.M.; Kumar, R.; Caldararu, O.; Peterson, K.; Wernersson, S.; Brath, U.; Leffler, H.; Oksanen, E.; et al. Entropy–Entropy Compensation between the Protein, Ligand, and Solvent Degrees of Freedom Fine-Tunes Affinity in Ligand Binding to Galectin-3C. JACS Au 2021, 1, 484-500, doi:10.1021/jacsau.0c00094.
15. Kozielski, F.; Sele, C.; Talibov, V.O.; Lou, J.; Dong, D.; Wang, Q.; Shi, X.; Nyblom, M.; Rogstam, A.; Krojer, T.; et al. Identification of fragments binding to SARS-CoV-2 nsp10 reveals ligand-binding sites in conserved interfaces between nsp10 and nsp14/nsp16. RSC Chemical Biology 2021, doi:10.1039/D1CB00135C.

In 2020, we started to also ask our user for deposition of data at the Protein Data Bank (PDB) with relation to LP3 and for 2021 received these 5 PDB entries: 7PGE; 7ORR; 7ORU; 7ORV; 7ORW

Collaboration

We collaborate in various ways with wider society and organizations within and outside LU. In 2021 LP3 hosted 4 visitors. LP3s efforts for visibility, access and outreach are described in the equally named chapter above. LP3s role as a Gateway environment and collaborations with ESS, MAX IV, but also other Swedish Universities that led to the formation of PPS is described in the introduction of this report. LP3 participated in an application within the thematic collaborative initiative call of LU addressing pandemic spread with the title PandA (Pandemics and Alertness), which will in 2022 be incorporated as a stand alone theme into the new Lund University Virus Center.