Gerald Geroldinger1, Matthias Tonner1, Judith Quirgst1, Martin Walter2, Sritama De

Overview

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Key numbers

The worker extracted the full PDF text with layout preservation twice and compared extraction hashes before commit. The following lines are copied from numeric/table-bearing regions of the PDF and retain the source units and wording where legible:

plants (1). The content of Art in those plants, which can be as much as 2% of their dry
with 5 mg/L hemin, 25,000 U/L penicillin, and 25 mg/L streptomycin in 50 mL TubeSpin®
was diluted 1:10 to achieve OD600 nm values in the linear range (0 - 1 OD600 nm). The cell-
cultivated in DMEM (high glucose) with 1.5 g/L NaHCO3, 25,000 U/L penicillin, 25 mg/L
suspension 1:1 with a formaldehyde solution (2%, v/v), aliquots of the resulting suspension
A suspension of LtP in YEM/PBS (1:1 v/v, including antibiotics and 6 µM hemin or 10%
compound stocks (vehicle DMSO, max. 1% final concentration) were added and five 1:5
untreated LtP (100% activity) were also loaded. The plate was incubated at 26 °C for 48 h.
25,000 U/L penicillin, 25 mg/L streptomycin, and 10% FCS was prepared. Two hundred µL
remaining rows the cells were incubated for 24 h at 37 °C and 5% CO2 to allow attachment.
plates for another 24 h at 37 °C and 5% CO2, 50 µL of resazurin solution (final
with addition of 10% glycerol). Aliquots of 500 µL were then transferred to syringes (Braun
ppm, 0.05 ppm, 0.1 ppm, 0.5 ppm, 1 ppm, 5 ppm, 10 ppm. Since the Sn concentrations in
calculated from diluted standard (0.5 mol/L HNO3) Sn calibration samples (0.1 ppb, 0.5
ppb, 1 ppb, 5 ppb, 10 ppb, 50 ppb and 100 ppb) and corrected with the isotope abundancy of
(RP-18, 250 x 4 mm, 5 µm, Merck, Germany). Mobile phase A was ultrapure water with 5%
acetonitrile (ACN) and 0.05% trifluoroacetic acid. Mobile phase C was ACN with 0.05%
min. Then the amount of A was decreased to 25% whereas phase C was increased to 75%
over 40 min. Phase C was held at 75% for 20 min and then further increased to 100% over
10 min. Finally, phase C was kept at 100% for 70 min. For the whole elution the flow rate
ultrapure water containing 0.1% HCOOH, and solvent B was ACN containing 0.1%
HCOOH. The applied gradient was as follows: 0-10 min: 5% B; 10-50 min: 575% B; 50-70
detector (FID), was used. The column was an Elite-5 MS (5% diphenyl-, 95% dimethyl-
EM voltage 1224, mass range 40 - 400, MS source 230 °C, MS quad 150 °C. Initial oven
a four-parameter logistic model (36) and expressed as the mean ± standard deviation (SD).
derivatives showed IC50 values for the viability of LtP in the low micromolar range in
Based on these data for YEM/PBS with 6 µM hemin and YEM/PBS with 10% FCS total
detects the Fe3+ portion of the chelatable iron in the cell (LIP). In the presence of DFO all
a limited toxicity in LtP (SnMP: IC50 = 64.1 ± 21.3 µM, n = 4, CrMP: IC50 = 58.6 ± 16.4
µM, n = 3, ZnPP: IC50 = 5.21 ± 0.57, n = 4). Therefore, we used SnMP and CrMP in
in which the viability was compromised less than 15%. In contrast, PIH and NAC were less
detectable amounts of BR (Fig. 4D). Therefore, we used RLS as a source of BVR in the LtP
of RLS (66 μL/mL) yielded photometric absorptions equivalent to 69.7 ± 0.22 pmol BR (n =
0.45 pmol BR (n = 2) equivalents in 30 min. This unequivocally demonstrates the absence of
analysis revealed that P4 was Art and P3 was a thermal rearrangement product of Art (Fig.
because dithionite-derived SO2 •- radicals (aN = 14.46 G, aH = 15.74 G; DMPO/SO2’-,
(66%, DMPO/•CO acyl radical) and a second adduct with aN = 16.24 G / aH = 22.21 G
adducts of aN = 14.88 G / aH = 17.69 G (84%, DMPO/•CO acyl radical) and aN = 13.44 G /
(47%, DMPO/•CO acyl radical) and aN = 14.46 G / aH = 13.53 G (53%) were observed.
DMPO/•C) and an ascorbyl radical (aH = 1.70 G, 5%) were recorded. NADPH as reductant
DMPO/•CO acyl radical) and aN = 13.78 G / aH = 15.10 G (71%). In the absence of Art in
this experiment an adduct with aN = 13.88 G / aH = 14.86 G (100%) was observed. Although

Methods (brief)

coupled plasma methods (ICP-OES, ICP-MS) we showed that these inhibitors do not block iron
(pH 7.4)/glucose (15 mM) solution. Samples were then incubated for 1 h at 26 °C under
For measurement the frozen samples were transferred to a quartz finger dewar and then
complex was performed as described previously (12). The LtP samples had protein
then collected in an 1.5 mL reaction tube and frozen to 77 K for analysis. Per incubation
Afterwards, the homogenized suspension was pipetted into digestion flasks and incubated
for 16 h. Then, the samples were heated to 75 °C for 2 h to complete the digestion.
spectrometry (ICP-OES) measurements. ICP-OES measurements were made using an
multielement analysis. The metal content of samples was calculated from diluted standard
(1.4 mol/L HNO3) calibration samples with the following multielement concentrations: 0.01
most samples were too low for detection by ICP-OES, aliquots of the digestion solution
spectrometry (ICP-MS) (Agilent 7700x). The content of 120Sn in the digestion solutions was
calculated from diluted standard (0.5 mol/L HNO3) Sn calibration samples (0.1 ppb, 0.5
used for LtP cultures were measured by ICP-OES after 10-fold dilution and acidification
calibration samples. However, BHI and FCS were acidified with concentrated HNO3
concentration of 1.14 mol/L by ICP-OES using diluted standard (1.14 mol/L HNO3)
Five mL of LtP suspension (1965 × 106 LtP) per sample were centrifuged at 3000 × g for 10
500 µM Art were added. The samples were then incubated in the dark for 30 min at 27 °C.

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Verification notes

Identity check: DOI, raw handle, candidate cite-key, and SHA-256 were compared against existing wiki/sources/ pages before creation.
Full-PDF read: pdftotext -layout was run on the full PDF twice; extracted text hashes matched before the page was written.
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Update history

The five most recent substantive edits to this page, classified major (evidence or structure moved), correction (a published value or statement was wrong and has been fixed), or minor (narrative rewritten without changing the underlying evidence). Each description is derived from what the edit did to this page; the linked commit is the authoritative record, routine regeneration passes are excluded, and the full version history lives in git. When DOI minting comes online (see schema docs), each entry below will also link to a version-pinned DataCite DOI.

Commit	Date	Change	Description
b01ec52c	2026-08-04	major	2 sections added
d49e450f	2026-08-03	major	5 sections added; narrative text revised