Sequential pH-driven dimerization and stabilization of the N-terminal domain enables rapid spider silk formation

Nina Kronqvist; Martins Otikovs; Volodymyr Chmyrov; Gefei Chen; Marlene Andersson; Kerstin Nordling; Michael Landreh; Médoune Sarr; Hans Jörnvall; Stefan Wennmalm; Jerker Widengren; Qing Meng; Anna Rising; Daniel Otzen; Stefan D Knight; Kristaps Jaudzems; Jan Johansson

doi:10.1038/ncomms4254

Sequential pH-driven dimerization and stabilization of the N-terminal domain enables rapid spider silk formation

Nina Kronqvist, Martins Otikovs, Volodymyr Chmyrov, Gefei Chen, Marlene Andersson, Kerstin Nordling, Michael Landreh, Médoune Sarr, Hans Jörnvall, Stefan Wennmalm, Jerker Widengren, Qing Meng, Anna Rising, Daniel Otzen, Stefan D Knight, Kristaps Jaudzems, Jan Johansson

Interdisciplinary Nanoscience Center - INANO-MBG, iNANO-huset

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

142 Citations (Scopus)

Abstract

The mechanisms controlling the conversion of spider silk proteins into insoluble fibres, which happens in a fraction of a second and in a defined region of the silk glands, are still unresolved. The N-terminal domain changes conformation and forms a homodimer when pH is lowered from 7 to 6; however, the molecular details still remain to be determined. Here we investigate site-directed mutants of the N-terminal domain from Euprosthenops australis major ampullate spidroin 1 and find that the charged residues D40, R60 and K65 mediate intersubunit electrostatic interactions. Protonation of E79 and E119 is required for structural conversions of the subunits into a dimer conformation, and subsequent protonation of E84 around pH 5.7 leads to the formation of a fully stable dimer. These residues are highly conserved, indicating that the now proposed three-step mechanism prevents premature aggregation of spidroins and enables fast formation of spider silk fibres in general.

Original language	English
Article number	3254
Journal	Nature Communications
Volume	5
Number of pages	11
ISSN	2041-1723
DOIs	https://doi.org/10.1038/ncomms4254
Publication status	Published - 10 Feb 2014

Access to Document

10.1038/ncomms4254

Cite this

Kronqvist, N., Otikovs, M., Chmyrov, V., Chen, G., Andersson, M., Nordling, K., Landreh, M., Sarr, M., Jörnvall, H., Wennmalm, S., Widengren, J., Meng, Q., Rising, A., Otzen, D., Knight, S. D., Jaudzems, K., & Johansson, J. (2014). Sequential pH-driven dimerization and stabilization of the N-terminal domain enables rapid spider silk formation. Nature Communications, 5, Article 3254. https://doi.org/10.1038/ncomms4254

@article{4c1f585e6bf646c5a0863792ed306300,

title = "Sequential pH-driven dimerization and stabilization of the N-terminal domain enables rapid spider silk formation",

abstract = "The mechanisms controlling the conversion of spider silk proteins into insoluble fibres, which happens in a fraction of a second and in a defined region of the silk glands, are still unresolved. The N-terminal domain changes conformation and forms a homodimer when pH is lowered from 7 to 6; however, the molecular details still remain to be determined. Here we investigate site-directed mutants of the N-terminal domain from Euprosthenops australis major ampullate spidroin 1 and find that the charged residues D40, R60 and K65 mediate intersubunit electrostatic interactions. Protonation of E79 and E119 is required for structural conversions of the subunits into a dimer conformation, and subsequent protonation of E84 around pH 5.7 leads to the formation of a fully stable dimer. These residues are highly conserved, indicating that the now proposed three-step mechanism prevents premature aggregation of spidroins and enables fast formation of spider silk fibres in general.",

author = "Nina Kronqvist and Martins Otikovs and Volodymyr Chmyrov and Gefei Chen and Marlene Andersson and Kerstin Nordling and Michael Landreh and M{\'e}doune Sarr and Hans J{\"o}rnvall and Stefan Wennmalm and Jerker Widengren and Qing Meng and Anna Rising and Daniel Otzen and Knight, {Stefan D} and Kristaps Jaudzems and Jan Johansson",

year = "2014",

month = feb,

day = "10",

doi = "10.1038/ncomms4254",

language = "English",

volume = "5",

journal = "Nature Communications",

issn = "2041-1723",

publisher = "Nature Research",

}

Kronqvist, N, Otikovs, M, Chmyrov, V, Chen, G, Andersson, M, Nordling, K, Landreh, M, Sarr, M, Jörnvall, H, Wennmalm, S, Widengren, J, Meng, Q, Rising, A, Otzen, D, Knight, SD, Jaudzems, K & Johansson, J 2014, 'Sequential pH-driven dimerization and stabilization of the N-terminal domain enables rapid spider silk formation', Nature Communications, vol. 5, 3254. https://doi.org/10.1038/ncomms4254

Sequential pH-driven dimerization and stabilization of the N-terminal domain enables rapid spider silk formation. / Kronqvist, Nina; Otikovs, Martins; Chmyrov, Volodymyr et al.
In: Nature Communications, Vol. 5, 3254, 10.02.2014.

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

TY - JOUR

T1 - Sequential pH-driven dimerization and stabilization of the N-terminal domain enables rapid spider silk formation

AU - Kronqvist, Nina

AU - Otikovs, Martins

AU - Chmyrov, Volodymyr

AU - Chen, Gefei

AU - Andersson, Marlene

AU - Nordling, Kerstin

AU - Landreh, Michael

AU - Sarr, Médoune

AU - Jörnvall, Hans

AU - Wennmalm, Stefan

AU - Widengren, Jerker

AU - Meng, Qing

AU - Rising, Anna

AU - Otzen, Daniel

AU - Knight, Stefan D

AU - Jaudzems, Kristaps

AU - Johansson, Jan

PY - 2014/2/10

Y1 - 2014/2/10

N2 - The mechanisms controlling the conversion of spider silk proteins into insoluble fibres, which happens in a fraction of a second and in a defined region of the silk glands, are still unresolved. The N-terminal domain changes conformation and forms a homodimer when pH is lowered from 7 to 6; however, the molecular details still remain to be determined. Here we investigate site-directed mutants of the N-terminal domain from Euprosthenops australis major ampullate spidroin 1 and find that the charged residues D40, R60 and K65 mediate intersubunit electrostatic interactions. Protonation of E79 and E119 is required for structural conversions of the subunits into a dimer conformation, and subsequent protonation of E84 around pH 5.7 leads to the formation of a fully stable dimer. These residues are highly conserved, indicating that the now proposed three-step mechanism prevents premature aggregation of spidroins and enables fast formation of spider silk fibres in general.

AB - The mechanisms controlling the conversion of spider silk proteins into insoluble fibres, which happens in a fraction of a second and in a defined region of the silk glands, are still unresolved. The N-terminal domain changes conformation and forms a homodimer when pH is lowered from 7 to 6; however, the molecular details still remain to be determined. Here we investigate site-directed mutants of the N-terminal domain from Euprosthenops australis major ampullate spidroin 1 and find that the charged residues D40, R60 and K65 mediate intersubunit electrostatic interactions. Protonation of E79 and E119 is required for structural conversions of the subunits into a dimer conformation, and subsequent protonation of E84 around pH 5.7 leads to the formation of a fully stable dimer. These residues are highly conserved, indicating that the now proposed three-step mechanism prevents premature aggregation of spidroins and enables fast formation of spider silk fibres in general.

U2 - 10.1038/ncomms4254

DO - 10.1038/ncomms4254

M3 - Journal article

C2 - 24510122

SN - 2041-1723

VL - 5

JO - Nature Communications

JF - Nature Communications

M1 - 3254

ER -

Sequential pH-driven dimerization and stabilization of the N-terminal domain enables rapid spider silk formation

Abstract

Access to Document

Fingerprint

Cite this