Steric regulation of tandem calponin homology domain actin-binding affinity
Journal Article
·
· Molecular Biology of the Cell
- Univ. of California, Berkeley, CA (United States)
- Department of Mechanical Engineering, University of California, Berkeley, Berkeley, CA 94720
- Lehrstuhl für Biophysik (E27), Technische Universität München, Garching 85748, Germany
- Univ. of California, Berkeley, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Chan Zuckerberg Biohub, San Francisco, CA (United States)
Tandem calponin homology (CH1–CH2) domains are common actin-binding domains in proteins that interact with and organize the actin cytoskeleton. Despite regions of high sequence similarity, CH1–CH2 domains can have remarkably different actin-binding properties, with disease-associated point mutants known to increase as well as decrease affinity for F-actin. To investigate features that affect CH1–CH2 affinity for F-actin in cells and in vitro, we perturbed the utrophin actin-binding domain by making point mutations at the CH1–CH2 interface, replacing the linker domain, and adding a polyethylene glycol (PEG) polymer to CH2. Consistent with a previous model describing CH2 as a steric negative regulator of actin binding, we find that utrophin CH1–CH2 affinity is both increased and decreased by modifications that change the effective “openness” of CH1 and CH2 in solution. We also identified interface mutations that caused a large increase in affinity without changing solution “openness,” suggesting additional influences on affinity. Interestingly, we also observe nonuniform subcellular localization of utrophin CH1–CH2 that depends on the N-terminal flanking region but not on bulk affinity. Finally, these observations provide new insights into how small sequence changes, such as those found in diseases, can affect CH1–CH2 binding properties.
- Research Organization:
- Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
- Sponsoring Organization:
- USDOE Office of Science (SC), Basic Energy Sciences (BES). Scientific User Facilities Division (SUF)
- Grant/Contract Number:
- AC02-05CH11231
- OSTI ID:
- 1615283
- Journal Information:
- Molecular Biology of the Cell, Journal Name: Molecular Biology of the Cell Journal Issue: 26 Vol. 30; ISSN 1059-1524
- Publisher:
- American Society for Cell BiologyCopyright Statement
- Country of Publication:
- United States
- Language:
- English
Similar Records
Biased localization of actin binding proteins by actin filament conformation
Transgelin-1 (SM22α) interacts with actin stress fibers and podosomes in smooth muscle cells without using its actin binding site
When fat is not bad: the regulation of actin dynamics by phospholipid signaling molecules
Journal Article
·
Tue Nov 24 19:00:00 EST 2020
· Nature Communications
·
OSTI ID:1815971
Transgelin-1 (SM22α) interacts with actin stress fibers and podosomes in smooth muscle cells without using its actin binding site
Journal Article
·
Wed Nov 14 23:00:00 EST 2018
· Biochemical and Biophysical Research Communications
·
OSTI ID:23107770
When fat is not bad: the regulation of actin dynamics by phospholipid signaling molecules
Journal Article
·
Tue Dec 31 19:00:00 EST 2013
· Frontiers in Plant Science
·
OSTI ID:1628251