Peterhoff, David and Zellner, Hermann and Guldan, Harald and Merkl, Rainer and Sterner, Reinhard and Babinger, Patrick (2012) Dimerization determines substrate specificity of a bacterial prenyltransferase. Chembiochem : a European journal of chemical biology 13 (9), pp. 1297-1303.
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We have identified the native dimer interface of heptaprenylglyceryl phosphate synthase PcrB from the bacterium Bacillus subtilis and analyzed the significance of oligomer formation for stability and catalytic activity. Computational methods predicted two different surface regions of the PcrB protomer that could be responsible for dimer formation. These bona fide interfaces were assessed both in silico and experimentally by the introduction of amino acid substitutions that led to monomerization, and by incorporation of an unnatural amino acid to allow cross-linking of the two protomers. The results showed that, in contrast to previous assumptions, PcrB uses the same interface for dimerization as the homologous geranylgeranylglyceryl phosphate synthase from Archaea. Thermal unfolding demonstrated that the monomeric proteins are only slightly less stable than wild-type PcrB. However, activity assays showed that monomerization limits the length of accepted polyprenyl pyrophosphates to three isoprene units, whereas the native PcrB substrate contains seven isoprene entities. We provide a plausible hypothesis as to how dimerization determines substrate specificity of PcrB.
|Institutions:||Biology, Preclinical Medicine > Institut für Biophysik und physikalische Biochemie > Prof. Dr. Reinhard Sterner > Arbeitsgruppe PD Dr. Rainer Merkl|
|Subjects:||500 Science > 570 Life sciences|
000 Computer science, information & general works > 004 Computer science
|Refereed:||Yes, this version has been refereed|
|Created at the University of Regensburg:||Yes|
|Deposited On:||09 Jul 2012 14:17|
|Last Modified:||09 Jul 2012 14:17|