Virginia Tech
Domain-based Bioinformatics Analysis and Molecular Insights for the Autoregulatory Mechanism of Phafin2
Abstract
dc:description.abstractPhafin2, an adaptor protein, is involved in various cellular processes, such as apoptosis, autophagy, endosomal cargo transportation, and macropinocytosis. Two domains, namely, PH and FYVE, contribute to Phafin2's cell membrane binding. Phafin2 also contains a poly aspartic acid (polyD) motif in its C-terminal region that can specifically autoinhibit the PH domain binding to membrane phosphatidylinositol 3-phosphate (PtdIns3P). Firstly, the study investigated the domain-based evolutionary pattern of PH, FYVE, and polyD motif of Phafin2 among its orthologs and Phafin2- like proteins. Using different bioinformatics tools and resources, it was concluded that the polyD motif only evolved in Phafin2 and PH- or both PH-FYVE-containing proteins of animals, highlighting the association in cellular functions that might have evolved uniquely in animals. Moreover, PH domain-free FYVE-containing proteins lack polyD motifs. Secondly, intramolecular autoregulatory and membrane binding properties of Phafin2 were studied by employing liposome co-sedimentation assay, isothermal titration calorimetry, and nuclear magnetic resonance spectroscopy. The residues Gly38, Lys45, Leu45, Lys51, Ala52, and Arg53 of the PH domain form a positively charged binding pocket that can bind the negatively charged polyD motif. The mutated Phafin2 PH domain (K51A/R53C and R53C) was unable to bind to synthetic polyD peptides, establishing the significance of those residues for the interaction between the PH domain and polyD motif. Moreover, the study also concluded that Phafin2-mediated membrane binding is not curvature-dependent.
Degree
thesis:*- Name thesis:degree_name
- Master of Science
- Level thesis:degree_level
- masters
- Discipline thesis:degree_discipline
- Biological Sciences
- Department dc:contributor.department
- Biological Sciences
- Grantor dc:publisher
- Virginia Tech
- Year dc:date.issued
- 2024
Author and committee
dc:creator, dc:contributor.*- Author dc:creator
-
- Hasan, Mahmudul
- Chair dc:contributor.committeechair
-
- Capelluto, Daniel G.
- Committee members dc:contributor.committeemember
-
- Zhang, Chenming
- Klemba, Michael Wade
- Lazar, Maria Iuliana
Subjects
dc:subject × 5Rights
dc:rights- Statement dc:rights
-
- In Copyright
- Licence dc:rights.uri
- Language dc:language.iso
- en
Identifiers
dc:identifier.*- Dc Identifier Other
- vt_gsexam:41261
- OAI identifier oai:identifier
- oai:vtechworks.lib.vt.edu:10919/120963