Colorimetric Determination of Tungsten and Molybdenum in Biological Samples
Molybdenum (Mo) and tungsten (W) are elements that are utilized in biological systems. They are typically incorporated into the catalytic sites of enzymes coordinated to an organic pyranopterin cofactor; Mo may also be present in the form of a FeMo cofactor. While Mo is used by all branches of life,...
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Bio-protocol LLC
2025-03-01
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| author | Aaron Donaghy Gerrit Schut Nana Shao Farris Poole Michael Adams |
| author_facet | Aaron Donaghy Gerrit Schut Nana Shao Farris Poole Michael Adams |
| author_sort | Aaron Donaghy |
| collection | DOAJ |
| description | Molybdenum (Mo) and tungsten (W) are elements that are utilized in biological systems. They are typically incorporated into the catalytic sites of enzymes coordinated to an organic pyranopterin cofactor; Mo may also be present in the form of a FeMo cofactor. While Mo is used by all branches of life, only a few microbes are able to utilize W. In order to study Mo- and W-dependent enzymes, it is important to be able to measure Mo and W in biological samples. Methods for determining Mo and W content in biological samples currently involve expensive and time-consuming processes like inductively coupled plasma mass spectrometry (ICP-MS) and chelation ion chromatography. There are less intensive colorimetric methods for measuring W in abiotic samples, but these have not been adapted to biological samples like cytosolic extracts and purified proteins. Herein, we developed a colorimetric assay based on the complexation of quercetin to molybdate (MoO42-) or tungstate (WO42-), the oxyanion forms of Mo and W that readily form in denatured biological samples. In the assay, the absorbance of quercetin is redshifted proportionally to the concentration of tungsten or molybdenum, which can be measured spectrophotometrically. This protocol provides a rapid method for screening biological samples for both Mo and W, although it does not distinguish between them. |
| format | Article |
| id | doaj-art-fa9604fefb364a2d9f06e52af957550b |
| institution | DOAJ |
| issn | 2331-8325 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | Bio-protocol LLC |
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| spelling | doaj-art-fa9604fefb364a2d9f06e52af957550b2025-08-20T03:15:31ZengBio-protocol LLCBio-Protocol2331-83252025-03-0115510.21769/BioProtoc.5195Colorimetric Determination of Tungsten and Molybdenum in Biological SamplesAaron Donaghy0Gerrit Schut1Nana Shao2Farris Poole3Michael Adams4Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, USADepartment of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, USADepartment of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, USADepartment of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, USADepartment of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, USAMolybdenum (Mo) and tungsten (W) are elements that are utilized in biological systems. They are typically incorporated into the catalytic sites of enzymes coordinated to an organic pyranopterin cofactor; Mo may also be present in the form of a FeMo cofactor. While Mo is used by all branches of life, only a few microbes are able to utilize W. In order to study Mo- and W-dependent enzymes, it is important to be able to measure Mo and W in biological samples. Methods for determining Mo and W content in biological samples currently involve expensive and time-consuming processes like inductively coupled plasma mass spectrometry (ICP-MS) and chelation ion chromatography. There are less intensive colorimetric methods for measuring W in abiotic samples, but these have not been adapted to biological samples like cytosolic extracts and purified proteins. Herein, we developed a colorimetric assay based on the complexation of quercetin to molybdate (MoO42-) or tungstate (WO42-), the oxyanion forms of Mo and W that readily form in denatured biological samples. In the assay, the absorbance of quercetin is redshifted proportionally to the concentration of tungsten or molybdenum, which can be measured spectrophotometrically. This protocol provides a rapid method for screening biological samples for both Mo and W, although it does not distinguish between them.https://bio-protocol.org/en/bpdetail?id=5195&type=0 |
| spellingShingle | Aaron Donaghy Gerrit Schut Nana Shao Farris Poole Michael Adams Colorimetric Determination of Tungsten and Molybdenum in Biological Samples Bio-Protocol |
| title | Colorimetric Determination of Tungsten and Molybdenum in Biological Samples |
| title_full | Colorimetric Determination of Tungsten and Molybdenum in Biological Samples |
| title_fullStr | Colorimetric Determination of Tungsten and Molybdenum in Biological Samples |
| title_full_unstemmed | Colorimetric Determination of Tungsten and Molybdenum in Biological Samples |
| title_short | Colorimetric Determination of Tungsten and Molybdenum in Biological Samples |
| title_sort | colorimetric determination of tungsten and molybdenum in biological samples |
| url | https://bio-protocol.org/en/bpdetail?id=5195&type=0 |
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