We have been awarded a Bronze medal in the UCL LEAF scheme for our efforts to reduce waste, to recycle and improve lab efficiency in general. Thanks to all lab members who contributes and for Nicola in particular for organising our submission to LEAF.
Not sure if there was a proper consultation on this, but the 3D printer has a name now.
After 20+ years of determining atomic level structures of proteins in three dimensions, we are going properly 3D. Our Prusa i3 MKS3+ has arrived!
HPLC plays up with random UV peaks, fraction collector tries to fail its calibration, centrifuge bottles crack with half of mammalian expression culture lost, fluorescent detector cuts off randomly.
And it’s only a Wednesday.
Panton Arms, here we come.
Welcome to Sarah and Bocheng for joining the group for their part II projects (third year undergraduate project in normal terminology), under the supervision of Shiv and Emma, respectively.
BioRxiv, posted 12 Jan 2023
DOI: 10.1101/2023.01.10.523494
PDB coordinates: 8BE1 (3D view)
Plasmids in Addgene: pExp-His-ZBasic-RBD, pExp-His-ZBasic-RBD-Avi
Monoclonal antibodies are increasingly used to prevent and treat viral infections, playing a pivotal role in pandemic response efforts. Antibody secreting cells (ASCs, plasma cells and plasmablasts) are an excellent source of high-affinity antibodies with therapeutic potential. Current methodologies to study antigen-specific ASCs either have low throughput, require expensive and labour-intensive screening or are technically demanding and therefore not accessible to the wider research community. Here, we present a straightforward technology for the rapid discovery of monoclonal antibodies from ASCs: we combine microfluidic encapsulation of single cells into an antibody capture hydrogel with antigen bait sorting by conventional flow cytometry. Continue reading →
Communications Biology 6: 34 (2023)
DOI: 10.1038/s42003-022-04388-4
Pubmed: 36635368
TGFβs, BMPs and Activins regulate numerous developmental and homeostatic processes and signal through hetero-tetrameric receptor complexes composed of two types of serine/threonine kinase receptors. Each of the 33 different ligands possesses unique affinities towards specific receptor types. However, the lack of specific tools hampered simultaneous testing of ligand binding towards all BMP/TGFβ receptors. Here we present a N-terminally Halo- and SNAP-tagged TGFβ/BMP receptor library to visualize receptor complexes in dual color. In combination with fluorescently labeled ligands, we established a Ligand Surface Binding Assay (LSBA) for optical quantification of receptor-dependent ligand binding in a cellular context. We highlight that LSBA is generally applicable to test (i) binding of different ligands such as Activin A, TGFβ1 and BMP9, (ii) for mutant screens and (iii) evolutionary comparisons. This experimental set-up opens opportunities for visualizing ligand-receptor binding dynamics, essential to determine signaling specificity and is easily adaptable for other receptor signaling pathways.
ChemBioChem, accepted article 17th Oct (2022)
DOI: 0.1002/cbic.202200339
Pubmed: 36250581
Enzymes are effective biological catalysts that accelerate almost all metabolic reactions in living organisms. Synthetic modulators of enzymes are useful tools for the study of enzymatic reactions and can provide starting points for the design of new drugs. Here, we report on the discovery of a class of biologically active compounds that covalently modifies lysine residues in human liver pyruvate kinase (PKL), leading to allosteric activation of the enzyme (EC50 = 0.29 µM). Surprisingly, the allosteric activation control point resides on the lysine residue K282 present in the catalytic site of PKL. These findings were confirmed by structural data, MS/MS experiments, and molecular modelling studies. Altogether, our study provides a molecular basis for the activation mechanism and establishes a framework for further development of human liver pyruvate kinase covalent activators.
Acta Crystallographica section F 78: 348-353 (2022)
DOI: 10.1107/S2053230X22008767
PDB coordinates:
7z39 (3D view)
The small molecule belumosudil was initially identified as a selective inhibitor of Rho-associated coiled-coil kinase 2 (ROCK2) and has recently been approved for the treatment of graft-versus-host disease. However, recent studies have shown that many of the phenotypes displayed upon treatment with belumosudil were due to CK2α inhibition. CK2α is in itself a very promising therapeutic target for a range of conditions and has recently been put forward as a potential treatment for COVID-19. Belumosudil presents a promising starting point for the development of future CK2α inhibitors as it provides a safe, potent and orally bioavailable scaffold. Continue reading →
Journal volume: RSC Med. Chem., 2022, Advance Article
DOI: 10.1039/D2MD00161F
CK2 is a ubiquitous protein kinase with an anti-apoptotic role and is found to be overexpressed in multiple cancer types. To this end, the inhibition of CK2 is of great interest with regard to the development of novel anti-cancer therapeutics. ATP-site inhibition of CK2 is possible; however, this typically results in poor selectivity due to the highly conserved nature of the catalytic site amongst kinases. An alternative methodology for the modulation of CK2 activity is through allosteric inhibition. The recently identified αD site represents a promising binding site for allosteric inhibition of CK2α. The work presented herein describes the development of a series of CK2α allosteric inhibitors through iterative cycles of X-ray crystallography and enzymatic assays, in addition to both fragment growing and fragment merging design strategies. Continue reading →