Crystallization and preliminary X-ray diffraction analysis of monoprenylated Rab7 GTPase in complex with Rab escort protein 1.

Alexey Rak Olena Pylypenko 1 Anca Niculae Roger Goody Kirill Alexandrov
1 Motilité structurale
CDC - Compartimentation et dynamique cellulaires
Abstract : Posttranslational geranylgeranylation of Rab GTPases is catalyzed by Rab geranylgeranyltransferase (RabGGTase), which consists of a catalytic alpha/beta heterodimer and an accessory Rab escort protein (REP). REP functions as a molecular chaperone that presents Rab proteins to the RabGGTase and after prenylation delivers them to their target membrane. Mutations in the REP-1 gene in humans lead to an X-chromosome-linked defect known as choroideremia, a progressive disease that inevitably culminates in complete blindness. Here we report in vitro assembly, purification, and crystallization of the monoprenylated Rab7GDP:REP-1 complex. X-Ray diffraction data for the REP-1:Rab7 complex were collected to 2.2-A resolution at the ESRF. The crystals belong to the orthorhombic space group P2(1)2(1)2 with unit-cell parameters a=64.3A, b=105.3A, c=132.6A. Preliminary structural analysis revealed the presence of one complex in the asymmetric unit. To understand the conformational changes in Rab protein on complex formation we also crystallized the GDP-bound form of Rab7 that diffracted to at least 1.8A on the in-house X-ray source.
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Journal articles
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https://hal.archives-ouvertes.fr/hal-02349781
Contributor : Olena Pylypenko <>
Submitted on : Tuesday, November 5, 2019 - 6:18:40 PM
Last modification on : Thursday, November 7, 2019 - 1:42:39 AM

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  • HAL Id : hal-02349781, version 1
  • PUBMED : 12576024

Citation

Alexey Rak, Olena Pylypenko, Anca Niculae, Roger Goody, Kirill Alexandrov. Crystallization and preliminary X-ray diffraction analysis of monoprenylated Rab7 GTPase in complex with Rab escort protein 1.. Journal of Structural Biology, Elsevier, 2003, 141 (1), pp.93-5. ⟨hal-02349781⟩

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