The N-terminal domains of spider silk proteins assemble ultrafast and protected from charge screening.
Web spiders assemble spidroin monomers into silk fibres of unrivalled tensile strength at remarkably high spinning speeds of up to 1 m s(-1). The spidroin N-terminal domain contains a charge-driven, pH-sensitive relay that controls self-association by an elusive mechanism. The underlying kinetics have not yet been reported. Here we engineer a fluorescence switch into the isolated N-terminal domain from spidroin 1 of the major ampullate gland of the nursery web spider E. australis that monitors dimerization. We observe ultrafast association that is surprisingly insensitive to salt, contrasting the classical screening effects in accelerated, charged protein interfaces. To gain deeper mechanistic insight, we mutate each of the protonatable residue side chains and probe their contributions. Two vicinal aspartic acids are critically involved in an unusual process of accelerated protein association that is protected from screening by electrolytes, potentially facilitating the rapid synthesis of silk fibres by web spiders.
Department of Biotechnology and Biophysics, Julius-Maximilians-University Würzburg, Am Hubland, 97074 Würzburg, Germany.
Simone
S
Schwarze
Fabian U
FU
Zwettler
Christopher M
CM
Johnson
Hannes
H
Neuweiler
D000818:Animals; D005349:Fibroins / Q000737:chemistry* / Q000235:genetics; D005453:Fluorescence; D007700:Kinetics; D017434:Protein Structure, Tertiary / Q000235:genetics; D013112:Spiders
C086087:spidroin 1; D005349:Fibroins