Crystal structure of tetrameric human Rabin8 GEF domain

Melanie Vetter; Niels Boegholm; Anni Christensen; Sagar Bhogaraju; Marie B Andersen; Anna Lorentzen; Esben Lorentzen

doi:10.1002/prot.25455

Crystal structure of tetrameric human Rabin8 GEF domain

Melanie Vetter, Niels Boegholm, Anni Christensen, Sagar Bhogaraju, Marie B Andersen, Anna Lorentzen, Esben Lorentzen

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avis › Tidsskriftartikel › Forskning › peer review

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Abstract

Rab GTPases and their effectors, activators and guanine nucleotide exchange factors (GEFs) are essential for vesicular transport. Rab8 and its GEF Rabin8 function in formation of the cilium organelle important for developmental signaling and sensory reception. Here, we show by size exclusion chromatography and analytical ultracentrifugation that Rabin8 exists in equilibrium between dimers and tetramers. The crystal structure of tetrameric Rabin8 GEF domain reveals an occluded Rab8 binding site suggesting that this oligomer is enzymatically inactive, a notion we verify experimentally using Rabin8/Rab8 GEF assays. We outline a procedure for the purification of active dimeric Rabin8 GEF‐domain for in vitro activity assays.

Originalsprog	Engelsk
Tidsskrift	Proteins
Vol/bind	86
Nummer	4
Sider (fra-til)	405-413
Antal sider	9
ISSN	0887-3585
DOI	https://doi.org/10.1002/prot.25455
Status	Udgivet - 10 jan. 2018

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10.1002/prot.25455

Crystal structure og tetramericAccepteret manuskript, 2,33 MB

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abstract = "Rab GTPases and their effectors, activators and guanine nucleotide exchange factors (GEFs) are essential for vesicular transport. Rab8 and its GEF Rabin8 function in formation of the cilium organelle important for developmental signaling and sensory reception. Here, we show by size exclusion chromatography and analytical ultracentrifugation that Rabin8 exists in equilibrium between dimers and tetramers. The crystal structure of tetrameric Rabin8 GEF domain reveals an occluded Rab8 binding site suggesting that this oligomer is enzymatically inactive, a notion we verify experimentally using Rabin8/Rab8 GEF assays. We outline a procedure for the purification of active dimeric Rabin8 GEF‐domain for in vitro activity assays.",

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author = "Melanie Vetter and Niels Boegholm and Anni Christensen and Sagar Bhogaraju and Andersen, {Marie B} and Anna Lorentzen and Esben Lorentzen",

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T1 - Crystal structure of tetrameric human Rabin8 GEF domain

AU - Vetter, Melanie

AU - Boegholm, Niels

AU - Christensen, Anni

AU - Bhogaraju, Sagar

AU - Andersen, Marie B

AU - Lorentzen, Anna

AU - Lorentzen, Esben

PY - 2018/1/10

Y1 - 2018/1/10

N2 - Rab GTPases and their effectors, activators and guanine nucleotide exchange factors (GEFs) are essential for vesicular transport. Rab8 and its GEF Rabin8 function in formation of the cilium organelle important for developmental signaling and sensory reception. Here, we show by size exclusion chromatography and analytical ultracentrifugation that Rabin8 exists in equilibrium between dimers and tetramers. The crystal structure of tetrameric Rabin8 GEF domain reveals an occluded Rab8 binding site suggesting that this oligomer is enzymatically inactive, a notion we verify experimentally using Rabin8/Rab8 GEF assays. We outline a procedure for the purification of active dimeric Rabin8 GEF‐domain for in vitro activity assays.

AB - Rab GTPases and their effectors, activators and guanine nucleotide exchange factors (GEFs) are essential for vesicular transport. Rab8 and its GEF Rabin8 function in formation of the cilium organelle important for developmental signaling and sensory reception. Here, we show by size exclusion chromatography and analytical ultracentrifugation that Rabin8 exists in equilibrium between dimers and tetramers. The crystal structure of tetrameric Rabin8 GEF domain reveals an occluded Rab8 binding site suggesting that this oligomer is enzymatically inactive, a notion we verify experimentally using Rabin8/Rab8 GEF assays. We outline a procedure for the purification of active dimeric Rabin8 GEF‐domain for in vitro activity assays.

KW - Journal Article

U2 - 10.1002/prot.25455

DO - 10.1002/prot.25455

M3 - Journal article

C2 - 29318657

SN - 0887-3585

VL - 86

SP - 405

EP - 413

JO - Proteins

JF - Proteins

IS - 4

ER -

Crystal structure of tetrameric human Rabin8 GEF domain

Abstract

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