Exceptionally tight membrane-binding may explain the key role of the synaptotagmin-7 C2A domain in asynchronous neurotransmitter release

Voleti, Rashmi; Tomchick, Diana R.; Südhof, Thomas C.; Rizo, Josep

doi:10.1073/pnas.1710708114

Title: Exceptionally tight membrane-binding may explain the key role of the synaptotagmin-7 C₂A domain in asynchronous neurotransmitter release

Journal Article · Mon Sep 18 00:00:00 EDT 2017 · Proceedings of the National Academy of Sciences of the United States of America

DOI:https://doi.org/10.1073/pnas.1710708114· OSTI ID:1409103

Voleti, Rashmi ^[1]; Tomchick, Diana R. ^[1]; Südhof, Thomas C. ^[2]; Rizo, Josep ^[1]

Univ. of Texas Southwestern Medical Center, Dallas, TX (United States)
Stanford Univ. Medical School, Stanford, CA (United States)

Synaptotagmins (Syts) act as Ca²⁺ sensors in neurotransmitter release by virtue of Ca²⁺-binding to their two C₂ domains, but their mechanisms of action remain unclear. Puzzlingly, Ca²⁺-binding to the C₂B domain appears to dominate Syt1 function in synchronous release, whereas Ca²⁺-binding to the C₂A domain mediates Syt7 function in asynchronous release. Here we show that crystal structures of the Syt7 C₂A domain and C₂AB region, and analyses of intrinsic Ca²⁺-binding to the Syt7 C2 domains using isothermal titration calorimetry, did not reveal major differences that could explain functional differentiation between Syt7 and Syt1. However, using liposome titrations under Ca²⁺saturating conditions, we show that the Syt7 C₂A domain has a very high membrane affinity and dominates phospholipid binding to Syt7 in the presence or absence of l-α-phosphatidylinositol 4,5-diphosphate (PIP₂). For Syt1, the two Ca²⁺-saturated C₂ domains have similar affinities for membranes lacking PIP₂, but the C₂B domain dominates binding to PIP₂-containing membranes. Mutagenesis revealed that the dramatic differences in membrane affinity between the Syt1 and Syt7 C₂A domains arise in part from apparently conservative residue substitutions, showing how striking biochemical and functional differences can result from the cumulative effects of subtle residue substitutions. Viewed together, our results suggest that membrane affinity may be a key determinant of the functions of Syt C₂ domains in neurotransmitter release.

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Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER); National Institutes of Health (NIH); Welch Foundation

Grant/Contract Number:: AC02-06CH11357; S10OD018027; S10RR026461; I-1304; R35 NS097333; NS037200; NS049044

OSTI ID:: 1409103

Journal Information:: Proceedings of the National Academy of Sciences of the United States of America, Vol. 114, Issue 40; ISSN 0027-8424

Publisher:: National Academy of SciencesCopyright Statement

Country of Publication:: United States

Language:: ENGLISH

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Cited by: 25 works

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37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
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synaptotagmin-7
synaptotagmin-1
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X-ray crystallography
neurotransmitter release

Title: Exceptionally tight membrane-binding may explain the key role of the synaptotagmin-7 C2A domain in asynchronous neurotransmitter release

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Title: Exceptionally tight membrane-binding may explain the key role of the synaptotagmin-7 C₂A domain in asynchronous neurotransmitter release