Tools & Techniques

Alongside biological projects, my group develops and applies methods that make membrane-protein structural biology more tractable and interpretable, through a mix of in-house work and external collaborations. This includes engineered binders for crystallisation and cryo-EM, high-throughput expression and purification pipelines, biosensors for functional readouts, and mass-spectrometry approaches to protein dynamics such as HDX-MS (hydrogen-deuterium exchange) and carbene footprinting, which resolve conformational changes that static structures cannot capture.

Nanobodies and binders

Cryo-EM density of the SLC15A4 dimer with two bound nanobodies (purple) — Human SLC15A4 dimer (orange/teal) with two bound nanobodies (purple); OMass depositions PDB 8QSK, 8QSL, 8QSM, 8QSN.

Cryo-EM density of two SPNS2 molecules (purple) bridged head-to-head by a gluebody dimer GbD12 (salmon) — Gluebody-mediated SPNS2 (59 kDa each) dimer: two SPNS2 molecules (purple) bridged head-to-head by the gluebody GbD12 (salmon).

Two nanobodies, Nb B5 (orange) and Nb E8, bound to the gasdermin D membrane pore — Nanobodies Nb B5 (orange) and Nb E8 bound to the gasdermin D pore.

AlphaFold3 model of an anti-BRIL nanobody (blue, CDR loops highlighted) bound to the BRIL apocytochrome b562RIL helical bundle (light blue) — Anti-BRIL nanobody (blue, CDR loops highlighted) bound to BRIL helices H3 and H4 (light blue) — AlphaFold3 model. Click to rotate; cyan spheres mark the BRIL N/C termini (fusion attachment points).

Protein-binder toolbox for solute carriers (J Mol Biol, 2024) — 525 binders against 22 SLCs, validated with a cell-based pipeline (immunofluorescence, immunoprecipitation) and applied to structural studies on KCC3b (SLC12A6); cryo-EM maps EMD-50364, 50405, 50406, 50407.
Gluebodies: transferable nanobody mutations for reliable crystallisation (ACS Cent Sci, 2025)
Anti-gasdermin D nanobodies (Nb B5, Nb E8) that bind gasdermin D and prevent pore formation
Anti-BRIL nanobody — a nanobody against the BRIL (apocytochrome b562RIL) fusion domain, as an affinity reagent for purification of BRIL-tagged constructs
Di-Gembodies: covalent nanobody dimers for cryo-EM of small proteins (Nat Chem Biol, 2025)
Crystal structure of the Tspan15 LEL domain and its ADAM10 binding site (Structure, 2022) — enabled by Fab 1C12

Mass spectrometry for protein dynamics

We use HDX-MS and carbene footprinting to map ligand binding sites and conformational change, complementing static structural data from cryo-EM and X-ray crystallography.

Native MS and HDX-MS of the MC2R–MRAP complex and antagonist-driven conformational change (bioRxiv, 2026)
Multi-region carbene footprinting of the β1-adrenergic receptor (Chem Sci, 2025)
Carbene footprinting of Gasdermin D and Caspase-1 binding (JACS Au, 2023)

High-throughput expression and biochemistry

High-throughput expression and purification of human SLCs (JoVE, 2023)
Expression screening of integral membrane proteins using BacMam (Methods Mol Biol, 2020)

Target Enabling Packages (SGC)

Public Target Enabling Packages produced through the SGC Membrane Protein Structure & Function group, bundling reagents, assays and structural information for open use by the community.

SLC12A4 (KCC1) and SLC12A6 (KCC3) Target Enabling Package, v1 (2024) (PDF, thesgc.org)

Functional readouts and electrophysiology

Schematic of the GloRider BRET chloride biosensor: NLuc and chloride-sensitive E²GFP tethered to the membrane via a palmitoylation tag, with NLuc donor energy transferred to E²GFP — GloRider — a BRET chloride biosensor.

Acidification assay in Xenopus oocytes after 40 min, with a pH calibration ladder — Oocyte acidification assay to quantify proton export by the gastric proton pump.

BRET-based chloride biosensor for high-throughput KCC2 screening (SLAS Discov, 2025)
Cation transport measured by atomic absorption in Xenopus oocytes (JoVE, 2013)
Voltage-clamp fluorometry of the gastric H,K-ATPase (PLoS ONE, 2012)
Acidification assay for proton transport by the gastric H,K-ATPase in Xenopus oocytes (JBC, 2009)