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Title: A bacterial surface layer protein exploits multistep crystallization for rapid self-assembly

Abstract

Surface layers (S-layers) are crystalline protein coats surrounding microbial cells. S-layer proteins (SLPs) regulate their extracellular self-assembly by crystallizing when exposed to an environmental trigger. However, molecular mechanisms governing rapid protein crystallization in vivo or in vitro are largely unknown. Here, we demonstrate that the Caulobacter crescentusSLP readily crystallizes into sheets in vitro via a calcium-triggered multistep assembly pathway. This pathway involves 2 domains serving distinct functions in assembly. The C-terminal crystallization domain forms the physiological 2-dimensional (2D) crystal lattice, but full-length protein crystallizes multiple orders of magnitude faster due to the N-terminal nucleation domain. Observing crystallization using a time course of electron cryo-microscopy (Cryo-EM) imaging reveals a crystalline intermediate wherein N-terminal nucleation domains exhibit motional dynamics with respect to rigid lattice-forming crystallization domains. Dynamic flexibility between the 2 domains rationalizes efficient S-layer crystal nucleation on the curved cellular surface. Rate enhancement of protein crystallization by a discrete nucleation domain may enable engineering of kinetically controllable self-assembling 2D macromolecular nanomaterials.

Authors:
 [1];  [1];  [1];  [2];  [3];  [3];  [4];  [2];  [3]; ORCiD logo [2]; ORCiD logo [1]
  1. Stanford Univ., CA (United States); SLAC National Accelerator Lab., Menlo Park, CA (United States)
  2. Univ. of British Columbia, Vancouver, BC (Canada)
  3. SLAC National Accelerator Lab., Menlo Park, CA (United States)
  4. Stanford Univ., CA (United States)
Publication Date:
Research Org.:
SLAC National Accelerator Lab., Menlo Park, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
OSTI Identifier:
1579600
Alternate Identifier(s):
OSTI ID: 1591627
Grant/Contract Number:  
AC02-76SF00515; RGPIN 36574-11; 04802-15
Resource Type:
Published Article
Journal Name:
Proceedings of the National Academy of Sciences of the United States of America
Additional Journal Information:
Journal Volume: 117; Journal Issue: 1; Journal ID: ISSN 0027-8424
Publisher:
National Academy of Sciences
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; protein self-assembly; Cryo-EM time course; microbiology; biophysics; crystal nucleation

Citation Formats

Herrmann, Jonathan, Li, Po-Nan, Jabbarpour, Fatemeh, Chan, Anson C. K., Rajkovic, Ivan, Matsui, Tsutomu, Shapiro, Lucy, Smit, John, Weiss, Thomas M., Murphy, Michael E. P., and Wakatsuki, Soichi. A bacterial surface layer protein exploits multistep crystallization for rapid self-assembly. United States: N. p., 2019. Web. doi:10.1073/pnas.1909798116.
Herrmann, Jonathan, Li, Po-Nan, Jabbarpour, Fatemeh, Chan, Anson C. K., Rajkovic, Ivan, Matsui, Tsutomu, Shapiro, Lucy, Smit, John, Weiss, Thomas M., Murphy, Michael E. P., & Wakatsuki, Soichi. A bacterial surface layer protein exploits multistep crystallization for rapid self-assembly. United States. doi:10.1073/pnas.1909798116.
Herrmann, Jonathan, Li, Po-Nan, Jabbarpour, Fatemeh, Chan, Anson C. K., Rajkovic, Ivan, Matsui, Tsutomu, Shapiro, Lucy, Smit, John, Weiss, Thomas M., Murphy, Michael E. P., and Wakatsuki, Soichi. Tue . "A bacterial surface layer protein exploits multistep crystallization for rapid self-assembly". United States. doi:10.1073/pnas.1909798116.
@article{osti_1579600,
title = {A bacterial surface layer protein exploits multistep crystallization for rapid self-assembly},
author = {Herrmann, Jonathan and Li, Po-Nan and Jabbarpour, Fatemeh and Chan, Anson C. K. and Rajkovic, Ivan and Matsui, Tsutomu and Shapiro, Lucy and Smit, John and Weiss, Thomas M. and Murphy, Michael E. P. and Wakatsuki, Soichi},
abstractNote = {Surface layers (S-layers) are crystalline protein coats surrounding microbial cells. S-layer proteins (SLPs) regulate their extracellular self-assembly by crystallizing when exposed to an environmental trigger. However, molecular mechanisms governing rapid protein crystallization in vivo or in vitro are largely unknown. Here, we demonstrate that theCaulobacter crescentusSLP readily crystallizes into sheets in vitro via a calcium-triggered multistep assembly pathway. This pathway involves 2 domains serving distinct functions in assembly. The C-terminal crystallization domain forms the physiological 2-dimensional (2D) crystal lattice, but full-length protein crystallizes multiple orders of magnitude faster due to the N-terminal nucleation domain. Observing crystallization using a time course of electron cryo-microscopy (Cryo-EM) imaging reveals a crystalline intermediate wherein N-terminal nucleation domains exhibit motional dynamics with respect to rigid lattice-forming crystallization domains. Dynamic flexibility between the 2 domains rationalizes efficient S-layer crystal nucleation on the curved cellular surface. Rate enhancement of protein crystallization by a discrete nucleation domain may enable engineering of kinetically controllable self-assembling 2D macromolecular nanomaterials.},
doi = {10.1073/pnas.1909798116},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
number = 1,
volume = 117,
place = {United States},
year = {2019},
month = {12}
}

Journal Article:
Free Publicly Available Full Text
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DOI: 10.1073/pnas.1909798116

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