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Development of a reporter peptide that catalytically produces a fluorescent signal through α‐complementation
Author(s) -
Nishiyama Kotaro,
Ichihashi Norikazu,
Kazuta Yasuaki,
Yomo Tetsuya
Publication year - 2015
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1002/pro.2667
Subject(s) - complementation , bimolecular fluorescence complementation , protein fragment complementation assay , domain (mathematical analysis) , chemistry , fluorescence , peptide , signal peptide , terminal (telecommunication) , biochemistry , biophysics , biology , peptide sequence , phenotype , physics , gene , computer science , mathematical analysis , telecommunications , mathematics , quantum mechanics
In α‐complementation, inactive N‐terminal (α‐domain) and C‐terminal (ω‐domain) fragments of β‐galactosidase associate to reconstitute the active protein. To date, the effect of α‐domain size on α‐complementation activity has not been systematically investigated. In this study, we compared the complementation activities of α‐domains of various sizes using an in vitro system. We found that the complementation activities are similar for α‐domains comprising between 45 and 229 N‐terminal residues but are significantly decreased for those containing less than 37 residues. However, these smaller α‐domains (15 and 25 residues) exhibited sufficient α‐complementation activity for application as reporters.