A reporter system that discriminates EF‐hand‐sensor motifs from signal‐modulators at the single‐motif level
Copyright policy )A reporter system that discriminates EF‐hand‐sensor motifs from signal‐modulators at the single‐motif level
The T‐protein is a single‐polypeptide bi‐functional enzyme composed of a chorismate mutase domain fused to a prephenate dehydrogenase domain (TyrA). We replaced the chorismate mutase domain with canonical or pseudo‐Ca2+‐binding motifs (EF‐hand). Canonical‐EF‐hand‐motifs differentiate from pseudo‐EF‐hand‐motifs by experimenting a Ca2+‐dependent conformational change. The Ca2+‐free EF‐hand‐TyrA fusion‐proteins showed TyrA activity at the T‐protein level. Canonical‐EF‐hand‐TyrA fusions showed a Ca2+‐dependent loss of TyrA activity, but a pseudo‐EF‐hand‐TyrA fusion showed high TyrA activity level in excess‐Ca2+ conditions. Because TyrA activity exhibits robust changes in response to Ca2+‐dependent‐EF‐hand conformational alterations, TyrA could be a good Ca2+‐reporter enzyme. A chimeric canonical/pseudo‐EF‐hand strategy is proposed to confer pseudo‐EF‐hand motifs with a Ca2+‐dependent conformational change.
Prephenate Dehydrogenase, Site-directed mutagenesis, Monofunctional TyrA, Protein Conformation, Recombinant Fusion Proteins, Amino Acid Motifs, Molecular Sequence Data, Protein Structure, Tertiary, Amino Acid Substitution, Bacterial Proteins, Multienzyme Complexes, Escherichia coli, Calcium, Amino Acid Sequence, Protein design, Multidomain protein, Calcium-sensor system, Conserved Sequence
Prephenate Dehydrogenase, Site-directed mutagenesis, Monofunctional TyrA, Protein Conformation, Recombinant Fusion Proteins, Amino Acid Motifs, Molecular Sequence Data, Protein Structure, Tertiary, Amino Acid Substitution, Bacterial Proteins, Multienzyme Complexes, Escherichia coli, Calcium, Amino Acid Sequence, Protein design, Multidomain protein, Calcium-sensor system, Conserved Sequence
selected citations These citations are derived from selected sources.This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).2 popularity This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.Average influence This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).Average impulse This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.Average
Citations provided by BIP!Popularity provided by BIP!