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The hydroxylase component of soluble methane monooxygenase from Methylococcus capsulatus (Bath) exists in several forms as shown by electrospray-ionisation mass spectrometry
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UNSPECIFIED (1998) The hydroxylase component of soluble methane monooxygenase from Methylococcus capsulatus (Bath) exists in several forms as shown by electrospray-ionisation mass spectrometry. EUROPEAN JOURNAL OF BIOCHEMISTRY, 254 (3). pp. 602-609. ISSN 0014-2956.
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Abstract
The hydroxylase of the soluble methane monooxygenase from the bacterium Methylococcus capsulatus (Bath) has been investigated by means of electrospray-ionisation mass spectrometry (ESI-MS) and liquid chromatography ESI-MS (LC/ESI-MS), The hydroxylase is a non-heme diiron protein consisting of three pairs of non-identical subunits (alpha approximate to 60 kDa, beta approximate to 45 kDa and gamma approximate to 20 kDa). Liquid chromatographic separation of the hydroxylase subunits was required before MS analysis in order to detect the a-subunit. The masses measured for the three subunits were found to disagree with those calculated from their gene sequences, Experiments involving the use of CNBr and trypsin cleavage followed by LC/ESI-MS and MS/MS analyses permitted the location and correction of errors in the sequences deduced from the use of cDNA. The ESI-MS results also showed that the alpha-subunit of the hydroxylase exists in multiple forms which result from cleavage of the protein. This observation explains a number of enigmatic features of the protein previously reported in the literature and illustrates the pivotal role of ESI-MS in complementing data obtained from molecular biology for the characterisation of the primary sequence of proteins.
Item Type: | Journal Article | ||||
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Subjects: | Q Science > QD Chemistry | ||||
Journal or Publication Title: | EUROPEAN JOURNAL OF BIOCHEMISTRY | ||||
Publisher: | SPRINGER VERLAG | ||||
ISSN: | 0014-2956 | ||||
Official Date: | 15 June 1998 | ||||
Dates: |
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Volume: | 254 | ||||
Number: | 3 | ||||
Number of Pages: | 8 | ||||
Page Range: | pp. 602-609 | ||||
Publication Status: | Published |
Data sourced from Thomson Reuters' Web of Knowledge
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