Molecular characterization of uptake hydrogenase in Frankia
2005 (English)In: Biochemical Society Transactions, ISSN 0300-5127, E-ISSN 1470-8752, Vol. 33, no 1, 64-66 p.Article in journal (Refereed) Published
A molecular characterization of uptake hydrogenase in Fronkia was performed by using two-dimensional gel electrophoresis, matrix-assisted laser-desorption ionization-time-of-flight mass spectrometry, PCR amplification and Southern blotting. A polypeptide of approx. 60 kDa was recognized in Frankia UGL011102, AVCI1 and KBS on the two-dimensional gel by blotting with Ralstonia eutropha (Hox G) antibody. Further analysis by MS resulted in a peptide 'fingerprint', which was similar to the membrane-bound hydrogenase 2 large subunit (HYD2) in Escherichia coli. in addition, a 127 bp PCR fragment could also be amplified from Frankia AVCI1, which gave a 76% similarity with the large subunit of hydrogenase in, e.g., Azotobacter chrococcum, Bradyrhizobium japonicum and Rhizobium leguminosorum. Although immunological similarity between the small subunit of Frankia hydrogenase and that of other organisms has not yet been found, a PCR product of 500 bp could be amplified from the local source of Fronkia, the analysis of which gave 69 and 67% identity with the small subunit of hydrogenases in B. japonicum and R. leguminosorum respectively. A Southern-blot analysis further indicated evidence for the presence of the small hydrogenase subunit in other Fronkia strains, i.e. KBS, Avcl1 and Ccl3.
Place, publisher, year, edition, pages
London: Biochemical Society , 2005. Vol. 33, no 1, 64-66 p.
Amino Acid Sequence, Base Sequence, Blotting; Southern, DNA Primers, Electrophoresis; Gel; Two-Dimensional, Frankia/*enzymology, Hydrogenase/chemistry/*metabolism, Molecular Sequence Data, Sequence Homology; Amino Acid
IdentifiersURN: urn:nbn:se:umu:diva-2782PubMedID: 15667266OAI: oai:DiVA.org:umu-2782DiVA: diva2:141051