title: Specific labeling of streptavidin for better understanding of ligand modification in modular method for affinity labeling (MoAL).
abstract: We studied the specific labeling of streptavidin using the modular method for affinity labeling (MoAL) that we developed based on a catalytic amide-forming reaction using 2-chloro-4,6-dimethoxy-1,3,5-triazine (CDMT) and a tertiary amine catalyst. The primary structures of avidin and streptavidin are significantly different from each other, and streptavidin does not possess an acidic amino acid equivalent to Asp108 of avidin, which is the target acidic amino acid that was labeled using MoAL. However, using biotinylated modular ligand catalysts (MLC) originally designed for labeling avidin, the labeling of streptavidin was found to successfully proceed at Glu51, which is located in a different region. The present study indicates that MoAL is readily applicable to protein labeling without a precise design for MLC. The most important factor for the design of MLC is to ensure that the linker is of sufficient length to connect the ligands to a catalytic site.
authors.affiliation: Faculty of Pharmaceutical Sciences, Institute of Medical, Pharmaceutical, and Health Sciences, Kanazawa University.
authors.forename: Munetaka
authors.initials: M
authors.lastname: Kunishima
authors.forename: Daiki
authors.initials: D
authors.lastname: Kato
authors.forename: Shuichi
authors.initials: S
authors.lastname: Nakanishi
authors.forename: Masanori
authors.initials: M
authors.lastname: Kitamura
authors.forename: Kohei
authors.initials: K
authors.lastname: Yamada
authors.forename: Keiji
authors.initials: K
authors.lastname: Terao
authors.forename: Tomoya
authors.initials: T
authors.lastname: Asano
mesh_terms: D000345:Affinity Labels / Q000737:chemistry; D000577:Amides / Q000737:chemistry*; D000595:Amino Acid Sequence; D001360:Avidin / Q000737:chemistry; D019921:Biotinylation; D008024:Ligands; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D019809:Streptavidin / Q000737:chemistry*
chemical_list: D000345:Affinity Labels; D000577:Amides; D008024:Ligands; D001360:Avidin; D019809:Streptavidin