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Example sentences for: midasin
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However, such primordial di-protomeric AAA polypeptides would have to have been simpler than those present in NSF, ClpA and p97 and other AAA proteins of the current era, for the latter assemble into two-tiered hexameric assemblies, with the axis joining the two protomers in each polypeptide oriented perpendicular to the plane of the hexameric ring [ 14 ] . We cannot, at present, exclude an alternative model for midasin structure in which the hexameric AAA ring assembly is a dimeric two-layered structure formed by two midasin polypeptides arranged laterally with all six odd-numbered protomers in one layer and all six even-numbered protomers in the other.
In all the available organisms, the four central protomers of the midasin polypeptide, AAA2, AAA3, AAA4 and AAA5, possess canonical Walker A and B motifs, as well as the critical Asn in sensor 1 and Arg in sensor 2 that are required for proper function in other AAA proteins [ 15 ] . The presence of these critical residues, taken together with their higher level of average sequence conservation (Table 1), suggests that the four central protomers all retain an active enzymatic function.
The results demonstrated that the tagged midasin localizes predominantly to the nucleus (Fig.
However, the recent increased availability of high sensitivity mass-spectrometry for peptide identification greatly lessens the difficulty of detecting midasin in semi-purified cell fractions.
The 3'-end of the coding region of the midasin gene was fused to a tag encoding 3× hemagglutamin (HA) by using the PCR-mediated cassette procedure of Longtine and colleagues [ 60 ] . The PCR primers used to generate the cassette were 5'-ACTGATTTTG CGTCAATACT TTACAGACCT GGCATCCAGC CGGATCCCCG GGTTAATTAA-3' and 5'-TCGTGTAGTA AACCTCCTCT TCTTGGTTTT CACGATATAC GAATTCGAGC TCGTTTAAAC-3'.
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