Metalloproteome Factory: From Crude Extract to Identification of Metalloproteins via 3-Dimensional HPLC-ICP-MS
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F60077344%3A_____%2F25%3A00646246" target="_blank" >RIV/60077344:_____/25:00646246 - isvavai.cz</a>
Alternative codes found
RIV/60076658:12310/25:43909982
Result on the web
<a href="https://pubs.acs.org/doi/10.1021/acs.analchem.5c02775" target="_blank" >https://pubs.acs.org/doi/10.1021/acs.analchem.5c02775</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1021/acs.analchem.5c02775" target="_blank" >10.1021/acs.analchem.5c02775</a>
Alternative languages
Result language
angličtina
Original language name
Metalloproteome Factory: From Crude Extract to Identification of Metalloproteins via 3-Dimensional HPLC-ICP-MS
Original language description
Metalloproteins are essential for a vast array of biological processes in all living organisms. Identification of multiple metalloproteins from native expression (metalloproteomics) based on a single chromatography has low resolution and/or sensitivity, while classical protein purification lacks the versatility required for purifying multiple proteins for proteomics. We present a new metalloproteomics method based on three dimensions of metal-free liquid chromatography (HPLC) hyphenated online with inductively coupled sector-field mass spectrometry (ICP-sfMS): anion exchange chromatography (AEC) followed by either chromatofocusing (CF) or hydrophobic interaction chromatography (HIC) and finally size exclusion chromatography (SEC). We used this 3D technique to purify naturally abundant metalloproteins from crude extracts, mainly from plants but also from bacteria, to a quantity and purity sufficient for protein identification and quantification by mass spectrometry (Tims-TOF-MS). Protein mass spectrometry data are available via ProteomeXchange with identifier PXD056024. Sensitivity and signal/background ratio were sufficient to identify proteins at concentrations <1 nM and total amounts <1 pmol in the final SEC. As shown by application examples, the method can be immediately applied to various biological materials from a large range of organisms without individual optimization for single proteins or organisms. It can thus be used to resolve mechanisms of metal metabolism in various contexts, e.g., metal deficiency, nutrition, toxicity, and detoxification.
Czech name
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Czech description
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Classification
Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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OECD FORD branch
10608 - Biochemistry and molecular biology
Result continuities
Project
<a href="/en/project/EF15_003%2F0000336" target="_blank" >EF15_003/0000336: Metals, plants and people</a><br>
Continuities
I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace
Others
Publication year
2025
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Analytical Chemistry
ISSN
0003-2700
e-ISSN
1520-6882
Volume of the periodical
97
Issue of the periodical within the volume
32
Country of publishing house
US - UNITED STATES
Number of pages
9
Pages from-to
17641-17649
UT code for WoS article
001546156700001
EID of the result in the Scopus database
2-s2.0-105013683395