Mass spectrometry
Native nano-DESI images intact kidney proteins
Open access · cc by · source: Europe PMC
Ammonium-acetate nano-DESI mass spectrometry imaging maps folded proteins and a noncovalent S100-A6 dimer in rat kidney at finer spatial scale than LESA.
Study at a glance
- Design
- Animal / in-vitro — Native nano-DESI MS imaging of proteins/complexes on rat kidney sections
- N
- 10 μm rat kidney sections; imaging method development — no cohort N
- Population
- Rat kidney tissue sections and protein standards
- Outcome
- Spatial detection of native proteins and noncovalent complexes
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Key findings
Native nano-DESI resolved kidney anatomy more finely than prior LESA imaging (typical LESA spots ~600 μm). They identified proteins not seen by native LESA, including the noncovalent S100-A6 homodimer, ALBP, and PEBP1. The largest species detected was the RidA trimer at 42.6 kDa. Images took about 7 h per section.
Methodology
The authors built a custom nano-DESI source on an Orbitrap Eclipse and sampled protein standards plus 10 μm rat kidney sections from ammonium acetate with a small amount of C8E4 detergent. They acquired ion images at 20 μm/s with 200 μm line spacing, then used location-targeted tandem MS and proton-transfer charge reduction to identify native proteins in situ.
Limitations
The work is method development on one rat kidney, not a biological hypothesis test. Protein signals are weak versus lipids, so higher spatial resolution would likely cost intensity. PTCR and oversampling were not fully validated as native-protein imaging strategies.
How this study connects
Role on claims
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Evidence for the claim as stated.
Ammonium-acetate nano-DESI mass spectrometry imaging maps folded proteins and a noncovalent S100-A6 dimer in rat kidney at finer spatial scale than LESA.
Evidence for the claim as stated.
Soft ionisation preserves complexes that harsh ionisation destroys; imaging spatial resolution and native versus denaturing conditions are different experiments.
Evidence for the claim as stated.
Open questions
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Soft ionisation preserves complexes that harsh ionisation destroys; imaging spatial resolution and native versus denaturing conditions are different experiments.
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