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Effects of N ‐Glycosylation Deletions on Cypridina Luciferase Activity
Author(s) -
Yasuno Rie,
Mitani Yasuo,
Ohmiya Yoshihiro
Publication year - 2017
Publication title -
photochemistry and photobiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.818
H-Index - 131
eISSN - 1751-1097
pISSN - 0031-8655
DOI - 10.1111/php.12847
Subject(s) - glycosylation , thermostability , luciferase , mutant , secretion , biochemistry , chemistry , glycan , residue (chemistry) , n linked glycosylation , consensus sequence , peptide sequence , biology , glycoprotein , enzyme , gene , transfection
Cypridina luciferase (Cluc), a secreted luminescent protein identified from Cypridina noctiluca , has two N ‐glycosylation sites. In this study, we evaluated the effects of N ‐glycosylation on Cluc properties by creating site‐directed mutagenic modifications at the consensus sequence for N ‐glycosylation (Asn‐X‐Ser/Thr). Eight variants consisting of four single‐ and double‐residue mutants each were characterized. The producibility and relative specific activity were apparently reduced in mutant Cluc although the thermostability and secretion efficiency were not affected. These results suggested that N ‐glycosylation modifications and the proper amino acid sequence of the N ‐glycan binding sites of Cluc are required for the complete protein folding to form a stable catalytic center, for the proper conformation of substrate–protein interaction residues, or for both and that defects in the glycosylation modification are not related to secretion process and stability of the protein.