scholarly journals Strength, Not Depth: An Exploration of Differential Membrane Binding Kinetics of Synaptotagmin-1 and Synaptotagmin-7 C2 Domains

2015 ◽  
Vol 108 (2) ◽  
pp. 555a
Author(s):  
Joshua V. Vermaas ◽  
Emad Tajkhorshid
Keyword(s):  
Membrane Binding ◽  
Binding Kinetics ◽  
C2 Domains ◽  
Synaptotagmin 1 ◽  
Kinetics Of

scholarly journals Membrane Binding Kinetics of Endophilin N-BAR Domain

2012 ◽  
Vol 102 (3) ◽  
pp. 97a-98a
Author(s):  
Wan-Ting Hsieh ◽  
Tobias Baumgart
Keyword(s):  
Membrane Binding ◽  
Binding Kinetics ◽  
Bar Domain ◽  
Kinetics Of

scholarly journals Stepwise membrane binding of the C2 domains of the extended synaptotagmins revealed by optical tweezers

2021 ◽  
Author(s):  
Jinghua Ge ◽  
Xin Bian ◽  
Lu Ma ◽  
Yiying Cai ◽  
Yanghui Li ◽  
...  
Keyword(s):  
Optical Tweezers ◽  
Membrane Binding ◽  
Binding Energies ◽  
Dependent Manner ◽  
C2 Domains ◽  
Binding Domains ◽  
Lipid Exchange ◽  
Membrane Contacts ◽  
Kinetics Of

Abstract Extended synaptotagmins (E-Syts) mediate lipid exchange between the endoplasmic reticulum (ER) and the plasma membrane (PM). Anchored on ER, E-Syts bind the PM via an array of C2 domains in a Ca2+- and lipid-dependent manner, drawing the two membranes close to facilitate lipid exchange. How these C2 domains bind the PM and regulate the ER-PM distance have not been well understood. Here, we applied optical tweezers to dissect PM membrane binding by E-Syt1 and E-Syt2. We detected Ca2+- and lipid-dependent membrane binding kinetics of both E-Syts and determined the binding energies and rates of individual C2 domains or pairs. We incorporated these parameters in a theoretical model to recapitulate various properties of E-Syt-mediated membrane contacts observed in vivo, including their equilibrium distances and probabilities. Our methods can be applied to study other proteins containing multiple membrane-binding domains linked by disordered polypeptides.

scholarly journals Comparison of the membrane binding kinetics of bovine prothrombin and its fragment 1.

1993 ◽  
Vol 268 (31) ◽  
pp. 22984-22991
Author(s):  
K.H. Pearce ◽  
M Hof ◽  
B.R. Lentz ◽  
N.L. Thompson
Keyword(s):  
Membrane Binding ◽  
Binding Kinetics ◽  
Kinetics Of

scholarly journals Synaptotagmin-1 C2B domains cooperatively stabilize the fusion stalk via a master-servant mechanism

2021 ◽  
Author(s):  
Ary Lautaro Di Bartolo ◽  
Diego Masone
Keyword(s):  
Free Energy ◽  
Membrane Fusion ◽  
Membrane Binding ◽  
Vesicle Fusion ◽  
Fusion Process ◽  
Total Work ◽  
C2 Domains ◽  
Domain Interactions ◽  
Synaptotagmin 1 ◽  
Data Point

Synaptotagmin-1 is a low-affinity Ca2+ sensor that triggers synchronous vesicle fusion. It contains two similar C2 domains (C2A and C2B) that cooperate in membrane binding, being the C2B domain the main responsible for the membrane fusion process due to its polybasic patch KRLKKKKTTIKK (321-332). In this work, a master-servant mechanism between two identical C2B domains is shown to control the formation of the fusion stalk. Two regions in C2B are essential for the process, the well-known polybasic patch and a recently described pair of arginines (398,399). The master domain shows strong PIP2 interactions with its polybasic patch and its pair of arginines. At the same time, the servant analogously cooperates with the master to reduce the total work to form the fusion stalk. The strategic mutation (T328E,T329E) in both master and servant domains disrupts the cooperative mechanism, drastically increasing the free energy needed to induce the fusion stalk, however with negligible effects on the master domain interactions with PIP2. These data point to a difference in the behavior of the servant domain, which is unable to sustain its PIP2 interactions neither through its polybasic patch nor through its pair of arginines, in the end losing its ability to assist the master in the formation of the fusion stalk.

scholarly journals Synaptotagmin-1 membrane binding is driven by the C2B domain and assisted cooperatively by the C2A domain

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Clémence Gruget ◽  
Oscar Bello ◽  
Jeff Coleman ◽  
Shyam S. Krishnakumar ◽  
Eric Perez ◽  
...  
Keyword(s):  
Neurotransmitter Release ◽  
Mutational Analysis ◽  
Membrane Binding ◽  
Surface Force ◽  
Membrane Interaction ◽  
C2 Domains ◽  
Binding Energetics ◽  
Synaptotagmin 1 ◽  
Site Binding ◽  
C2a Domain

Abstract Synaptotagmin interaction with anionic lipid (phosphatidylserine/phosphatidylinositol) containing membranes, both in the absence and presence of calcium ions (Ca2+), is critical to its central role in orchestrating neurotransmitter release. The molecular surfaces involved, namely the conserved polylysine motif in the C2B domain and Ca2+-binding aliphatic loops on both C2A and C2B domains, are known. Here we use surface force apparatus combined with systematic mutational analysis of the functional surfaces to directly measure Syt1-membrane interaction and fully map the site-binding energetics of Syt1 both in the absence and presence of Ca2+. By correlating energetics data with the molecular rearrangements measured during confinement, we find that both C2 domains cooperate in membrane binding, with the C2B domain functioning as the main energetic driver, and the C2A domain acting as a facilitator.

Membrane Binding Kinetics of Protein Kinase C βII Mediated by the C2 Domain†

Biochemistry ◽  
2001 ◽  
Vol 40 (44) ◽  
pp. 13216-13229 ◽  
Author(s):  
Eric A. Nalefski ◽  
Alexandra C. Newton
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Membrane Binding ◽  
Binding Kinetics ◽  
C2 Domain ◽  
Kinase C ◽  
Kinetics Of

scholarly journals Membrane binding kinetics of factor VIII indicate a complex binding process.

1993 ◽  
Vol 268 (12) ◽  
pp. 8815-8824
Author(s):  
C. Bardelle ◽  
B. Furie ◽  
B.C. Furie ◽  
G.E. Gilbert
Keyword(s):  
Factor Viii ◽  
Membrane Binding ◽  
Binding Kinetics ◽  
Binding Process ◽  
Kinetics Of

The membrane binding kinetics of full-length PKCα is determined by membrane lipid composition

2012 ◽  
Vol 1821 (11) ◽  
pp. 1434-1442 ◽  
Author(s):  
Ángel Pérez-Lara ◽  
Antonio L. Egea-Jiménez ◽  
Alessio Ausili ◽  
Senena Corbalán-García ◽  
Juan C. Gómez-Fernández
Keyword(s):  
Lipid Composition ◽  
Membrane Binding ◽  
Membrane Lipid ◽  
Full Length ◽  
Binding Kinetics ◽  
Membrane Lipid Composition ◽  
Kinetics Of

scholarly journals Cooperativity and Avidity in Membrane Binding by C2AB Tandem Domains of Synaptotagmins 1 and 7

2018 ◽  
Author(s):  
H. Tran ◽  
L. Anderson ◽  
J. Knight
Keyword(s):  
Hormone Secretion ◽  
Membrane Binding ◽  
Dissociation Rate ◽  
Dissociation Kinetics ◽  
Kinetic Measurements ◽  
C2 Domains ◽  
Synaptotagmin 1 ◽  
Fusion Pores ◽  
Fluorescence Kinetic ◽  
C2a Domain

AbstractSynaptotagmin-1 (Syt-1) and synaptotagmin-7 (Syt-7) contain analogous tandem C2 domains, C2A and C2B, which together serve as a Ca2+ sensor to bind membranes and promote the stabilization of exocytotic fusion pores. Functionally, Syt-1 triggers fast release of neurotransmitters, while Syt-7 is involved in lower-Ca2+ processes such as hormone secretion. Evidence suggests that Syt-1 C2 domains bind membranes cooperatively, penetrating farther into membranes as the C2AB tandem than as individual C2 domains. In contrast, we previously reported that the two C2 domains of Syt-7 bind membranes independently, based in part on measurements of their liposome dissociation kinetics. Here, we have investigated the effects of C2A-C2B interdomain cooperativity with Syt-1 and Syt-7 using directly comparable measurements. We report Ca2+ sensitivities, dissociation kinetics, and membrane insertion using liposomes approximating physiological lipid compositions. Equilibrium Ca2+ titrations confirm that the Syt-7 C2AB tandem has a greater Ca2+ sensitivity of membrane binding than either of its individual domains. Stopped-flow fluorescence kinetic measurements show that Syt-1 C2AB dissociates from liposome membranes much more slowly than either of its isolated C2 domains, suggesting that the two C2 domains of Syt-1 bind membranes cooperatively. In contrast, the dominant population of Syt-7 C2AB has a dissociation rate comparable to its C2A domain, indicating a lack of cooperativity, while only a small subpopulation dissociates at a slower rate. Measurements using an environment-sensitive fluorescent probe indicate that the Syt-7 C2B domain inserts more deeply into membranes as part of the C2AB tandem, similarly to Syt-1. Overall, these measurements are consistent with a model in which the structural linkage of C2A and C2B impacts the membrane-binding geometry of synaptotagmin C2B domains, but imparts little or no cooperativity to Syt-7 membrane binding and dissociation events that are dominated by its C2A domain.

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