Single-molecule force spectroscopy of protein-membrane interactions

ELife
Lu MaYongli Zhang

Abstract

Many biological processes rely on protein-membrane interactions in the presence of mechanical forces, yet high resolution methods to quantify such interactions are lacking. Here, we describe a single-molecule force spectroscopy approach to quantify membrane binding of C2 domains in Synaptotagmin-1 (Syt1) and Extended Synaptotagmin-2 (E-Syt2). Syts and E-Syts bind the plasma membrane via multiple C2 domains, bridging the plasma membrane with synaptic vesicles or endoplasmic reticulum to regulate membrane fusion or lipid exchange, respectively. In our approach, single proteins attached to membranes supported on silica beads are pulled by optical tweezers, allowing membrane binding and unbinding transitions to be measured with unprecedented spatiotemporal resolution. C2 domains from either protein resisted unbinding forces of 2-7 pN and had binding energies of 4-14 kBT per C2 domain. Regulation by bilayer composition or Ca2+ recapitulated known properties of both proteins. The method can be widely applied to study protein-membrane interactions.

References

Aug 7, 1990·Biochemistry·K A Dill
Sep 9, 1994·Science·C BustamanteS Smith
Jun 11, 2004·Annual Review of Biochemistry·Carlos BustamanteDavid Izhaky
Nov 15, 2005·Nature·Elio A AbbondanzieriSteven M Block
Dec 2, 2005·Nature·Harvey T McMahon, Jennifer L Gallop
Apr 11, 2006·The Journal of Membrane Biology·Norbert KucerkaJohn F Nagle
Apr 18, 2006·Biochimica Et Biophysica Acta·James H Hurley
Jun 6, 2006·Proceedings of the National Academy of Sciences of the United States of America·Jeffrey R MoffittCarlos Bustamante
Nov 2, 2006·The Journal of Biological Chemistry·Mats X AnderssonAnna Stina Sandelius
Mar 16, 2007·Proceedings of the National Academy of Sciences of the United States of America·Sang-Won MinThomas C Südhof
Jan 25, 2008·Nature Reviews. Molecular Cell Biology·Mark A Lemmon
Feb 16, 2008·Annual Review of Biochemistry·Edwin R Chapman
Jan 24, 2009·Science·Thomas C Südhof, James E Rothman
Jan 27, 2009·Biophysical Journal·Jefferson D Knight, Joseph J Falke
Feb 3, 2009·Biophysical Journal·Kerry L FusonAndres F Oberhauser
Oct 21, 2010·Biophysical Journal·Hirohide TakahashiJ Michael Edwardson
Jun 7, 2011·Nature Structural & Molecular Biology·Geert van den BogaartReinhard Jahn
Sep 18, 2012·Cell·Hongying ShenPietro De Camilli
Oct 2, 2012·The Review of Scientific Instruments·George SirinakisYongli Zhang
Dec 10, 2013·Biophysical Journal·Shyam S KrishnakumarJames E Rothman
Jan 21, 2014·Biochimica Et Biophysica Acta·Senena Corbalan-Garcia, Juan C Gómez-Fernández
Sep 10, 2014·Proceedings of the National Academy of Sciences of the United States of America·Jing WangJames E Rothman
Sep 19, 2014·Biochemical Society Transactions·Marina E MonteiroFábio Fernandes
Dec 3, 2014·Biochimica Et Biophysica Acta·Kazuya Tsujita, Toshiki Itoh
Dec 17, 2014·Nature Communications·Chao-Chen LinPeter Jomo Walla

❮ Previous
Next ❯

Citations

May 25, 2018·Annual Review of Biophysics·Axel T BrungerQiangjun Zhou
Jul 20, 2019·Nature Chemical Biology·Xin BianChenxiang Lin
Nov 9, 2019·Physical Review Letters·Andriy Goychuk, Erwin Frey
Jan 19, 2020·Biophysical Journal·Raya SorkinGijs J L Wuite
Feb 13, 2021·Science·Swathi SudhakarErik Schäffer
Feb 6, 2020·Langmuir : the ACS Journal of Surfaces and Colloids·Tina R MatinGavin M King
May 1, 2018·Trends in Cell Biology·Axel T BrungerYing Lai
Nov 21, 2019·Nano Letters·Swathi SudhakarErik Schäffer
Dec 18, 2021·Nature Chemical Biology·Jinghua GeYongli Zhang

❮ Previous
Next ❯

Methods Mentioned

BETA
protein folding
optical tweezers
Fluorescence
atomic force microscopy
gel filtration

Software Mentioned

MATLAB

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.