Academic Journal
Determinants of Rab5 interaction with the N terminus of early endosome antigen 1
| العنوان: | Determinants of Rab5 interaction with the N terminus of early endosome antigen 1 |
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| المؤلفون: | Merithew, Eric Lee, Stone, Craig, Sudharshan, Eathiraj, Lambright, David G. |
| المساهمون: | Program in Molecular Medicine, Graduate School of Biomedical Sciences |
| المصدر: | The Journal of biological chemistry ; 278 ; 10 ; 8494-500 |
| سنة النشر: | 2022 |
| المجموعة: | University of Massachusetts, Medical School: eScholarship@UMMS |
| مصطلحات موضوعية: | Amino Acid Sequence, Binding Sites, Membrane Proteins, Molecular Sequence Data, Mutagenesis, Site-Directed, Protein Binding, Sequence Homology, Amino Acid, Surface Plasmon Resonance, Vesicular Transport Proteins, Zinc Fingers, rab5 GTP-Binding Proteins, Life Sciences, Medicine and Health Sciences |
| الوصف: | The Rab5 effector early endosome antigen 1 (EEA1) is a parallel coiled coil homodimer with an N-terminal C(2)H(2) Zn(2+) finger and a C-terminal FYVE domain. Rab5 binds to independent sites at the N and C terminus of EEA1. To gain further insight into the structural determinants for endosome tethering and fusion, we have characterized the interaction of Rab5C with truncation and site-specific mutants of EEA1 using quantitative binding measurements. The results demonstrate that the C(2)H(2) Zn(2+) finger is both essential and sufficient for the N-terminal interaction with Rab5. Although the heptad repeat C-terminal to the C(2)H(2) Zn(2+) finger provides the driving force for stable homodimerization, it does not influence either the affinity or stoichiometry of Rab5 binding. Hydrophobic residues predicted to cluster on a common face of the C(2)H(2) Zn(2+) finger play a critical role in the interaction with Rab5. Although the homologous C(2)H(2) Zn(2+) finger of the Rab5 effector Rabenosyn binds to Rab5 with comparable affinity, the analogous C(2)H(2) Zn(2+) finger of the yeast homologue Vac1 shows no detectable interaction with Rab5, reflecting non-conservative substitutions of critical residues. Large changes in the intrinsic tryptophan fluorescence of Rab5 accompany binding to the C(2)H(2) Zn(2+) finger of EEA1. These observations can be explained by a mode of interaction in which a partially exposed tryptophan residue located at the interface between the switch I and II regions of Rab5 lies within a hydrophobic interface with a cluster of non-polar residues in the C(2)H(2) Zn(2+) finger of EEA1. |
| نوع الوثيقة: | article in journal/newspaper |
| اللغة: | English |
| Relation: | Link to article in PubMed; http://dx.doi.org/10.1074/jbc.M211514200; J Biol Chem. 2003 Mar 7;278(10):8494-500. Epub 2002 Dec 19. Link to article on publisher's site; 0021-9258 (Print); http://hdl.handle.net/20.500.14038/34202; https://escholarship.umassmed.edu/gsbs_sp/859; 670705; gsbs_sp/859 |
| DOI: | 10.1074/jbc.M211514200 |
| الاتاحة: | https://doi.org/10.1074/jbc.M211514200 https://hdl.handle.net/20.500.14038/34202 https://escholarship.umassmed.edu/gsbs_sp/859 |
| رقم الانضمام: | edsbas.B38D9EBA |
| قاعدة البيانات: | BASE |
| DOI: | 10.1074/jbc.M211514200 |
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