Crystal structure of the RIM2 C2A-domain at 1.4 Å resolution

Han Dai, Diana R Tomchick, Jesús García, Thomas C. Südhof, Mischa Machius, Jose Rizo-Rey

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

RIMs are large proteins that contain two C2-domains and are localized at presynaptic active zones, where neurotransmitters are released. RIMs play key roles in synaptic vesicle priming and regulation of presynaptic plasticity. A mutation in the RIM1 C2A-domain has been implicated in autosomal dominant cone-rod dystrophy (CORD7). The RIM C2A-domain does not contain the full complement of aspartate residues that commonly mediate Ca2+ binding at the top loops of C2-domains, and has been reported to interact with SNAP-25 and synaptotagmin 1, two proteins from the Ca2+-dependent membrane fusion machinery. Here we have used NMR spectroscopy and X-ray crystallography to analyze the structure and biochemical properties of the RIM2 C2A-domain, which is closely related to the RIM1 C2A-domain. We find that the RIM2 C2A-domain does not bind Ca2+. Moreover, little binding of the RIM2 C 2A-domain to SNAP-25 and to the C2-domains of synaptotagmin 1 was detected by NMR experiments, suggesting that as yet unidentified interactions of the RIM C2A-domain mediate its function. The crystal structure of the RIM2 C2A-domain using data to 1.4 Å resolution reveals a β-sandwich that resembles those observed for other C2-domains, but exhibits a unique dipolar distribution of electrostatic charges whereby one edge of the β-sandwich is highly positive and the other edge is highly negative. The location of the mutation site implicated in CORD7 at the bottom of the domain and the pattern of sequence conservation suggest that, in contrast to most C2-domains, the RIM C2A-domains may function through Ca2+-independent interactions involving their bottom face.

Original languageEnglish (US)
Pages (from-to)13533-13542
Number of pages10
JournalBiochemistry
Volume44
Issue number41
DOIs
StatePublished - Oct 18 2005

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Reaction injection molding
Crystal structure
Synaptotagmin I
Mutation
Membrane Fusion
Synaptic Vesicles
X ray crystallography
X Ray Crystallography
Static Electricity
Aspartic Acid
Nuclear magnetic resonance spectroscopy
Machinery
Plasticity
Neurotransmitter Agents
Cones
Electrostatics
Conservation
Proteins
Magnetic Resonance Spectroscopy
Fusion reactions

ASJC Scopus subject areas

  • Biochemistry

Cite this

Crystal structure of the RIM2 C2A-domain at 1.4 Å resolution. / Dai, Han; Tomchick, Diana R; García, Jesús; Südhof, Thomas C.; Machius, Mischa; Rizo-Rey, Jose.

In: Biochemistry, Vol. 44, No. 41, 18.10.2005, p. 13533-13542.

Research output: Contribution to journalArticle

Dai, H, Tomchick, DR, García, J, Südhof, TC, Machius, M & Rizo-Rey, J 2005, 'Crystal structure of the RIM2 C2A-domain at 1.4 Å resolution', Biochemistry, vol. 44, no. 41, pp. 13533-13542. https://doi.org/10.1021/bi0513608
Dai, Han ; Tomchick, Diana R ; García, Jesús ; Südhof, Thomas C. ; Machius, Mischa ; Rizo-Rey, Jose. / Crystal structure of the RIM2 C2A-domain at 1.4 Å resolution. In: Biochemistry. 2005 ; Vol. 44, No. 41. pp. 13533-13542.
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