Septin 8 is an interaction partner and in vitro substrate of MK5

• 作者: Alexey Shiryaev Gianina Dumitriu Sergiy Kostenko Ugo Moens
• 来源期刊: 世界生物化学杂志：英文版（电子版）
• 学科: 生物学
• 关键词: MITOGEN-ACTIVATED PROTEIN kinase-activated PROTEIN kinase-5 Fluorescence resonance energy transfer SEPTIN Phosphorylation SYNAPTOPHYSIN
• 年，卷（期）: 2012,（5）
• 页码范围: 98-109
• 页数: 12页
• 分类号: Q55

摘要

AIM:To identify novel substrates for the mitogen-activated protein kinase-activated protein kinase 5(MK5).METHODS:Yeast two-hybrid screening with MK5 as bait was used to identify novel possible interaction partners.The binding of putative partner was further examined by glutathione S-transferase(GST) pull-down,co-immunoprecipitation and fluorescence resonance energy transfer(FRET) analysis.In vitro kinase and peptide array assays were used to map MK5 phosphoacceptor sites on the new partner.Confocal microscopy was performed to study the subcellular localization of MK5 and its partners.RESULTS:Septin 8 was identified as a novel interaction partner for MK5 by yeast two-hybrid screening.This interaction was confirmed by GST pull-down,coimmunoprecipitation and FRET analysis.Septin 5,which can form a complex with septin 8,did not interact with MK5.Serine residues 242 and 271 on septin 8 were identified as in vitro MK5 phosphorylation sites.MK5 and septin 8 co-localized in the perinuclear area and in cell protrusions.Moreover,both proteins co-localized with vesicle marker synaptophysin.