Academic Journal

Purification and identification of novel Rab effectors using affinity chromatography

التفاصيل البيبلوغرافية
العنوان: Purification and identification of novel Rab effectors using affinity chromatography
المؤلفون: Christoforidis, S., Zerial, M.
سنة النشر: 2000
مصطلحات موضوعية: Animals, Brain Chemistry, Cattle, Chromatography, Affinity/*methods, Guanine Nucleotide Exchange Factors/isolation & purification, Ligands, Membrane Proteins/isolation & purification, Nerve Tissue Proteins/*isolation & purification, Protein Binding, Vesicular Transport Proteins, rab5 GTP-Binding Proteins/*metabolism
الوصف: Rab GTPases are central regulatory elements of the intracellular transport machinery of eukaryotic cells. To regulate vesicle docking and fusion as well as organelle dynamics Rab proteins interact with effector molecules in the GTP-bound active state. The identification of Rab effectors is, therefore, of primary importance for the mechanistic understanding of intracellular transport. Here we describe the experimental system we have developed to biochemically purify and identify effectors of the small GTPase Rab5. The method, which is based on an affinity chromatography procedure, results in the large-scale purification of Rab effectors in amounts sufficient for both their identification by microsequencing techniques and their functional characterization. In the case of Rab5, the procedure allows a comprehensive analysis of the downstream effectors and regulators of this GTPase. We expect this strategy to provide fundamental insights into the molecular mechanism of membrane transport but also to be applicable to several other GTPase-dependent biological functions. ; Methods
نوع الوثيقة: article in journal/newspaper
اللغة: English
تدمد: 1046-2023
Relation: http://olympias.lib.uoi.gr/jspui/handle/123456789/18888
الاتاحة: http://olympias.lib.uoi.gr/jspui/handle/123456789/18888
Rights: Default Licence ; campus
رقم الانضمام: edsbas.83295261
قاعدة البيانات: BASE