Independent Membrane Binding Properties of the Caspase Generated Fragments of the Beaded Filament Structural Protein 1 (BFSP1) Involves an Amphipathic Helix
Background: BFSP1 (beaded filament structural protein 1) is a plasma membrane, Aquaporin 0 (AQP0/MIP)-associated intermediate filament protein expressed in the eye lens. BFSP1 is myristoylated, a post-translation modification that requires caspase cleavage at D433. Bioinformatic analyses suggested t...
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| Format: | Article |
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MDPI AG
2023-06-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/12/12/1580 |
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| author | Miguel Jarrin Alexia A. Kalligeraki Alice Uwineza Chris S. Cawood Adrian P. Brown Edward N. Ward Khoa Le Stefanie Freitag-Pohl Ehmke Pohl Bence Kiss Antal Tapodi Roy A. Quinlan |
| author_facet | Miguel Jarrin Alexia A. Kalligeraki Alice Uwineza Chris S. Cawood Adrian P. Brown Edward N. Ward Khoa Le Stefanie Freitag-Pohl Ehmke Pohl Bence Kiss Antal Tapodi Roy A. Quinlan |
| author_sort | Miguel Jarrin |
| collection | DOAJ |
| description | Background: BFSP1 (beaded filament structural protein 1) is a plasma membrane, Aquaporin 0 (AQP0/MIP)-associated intermediate filament protein expressed in the eye lens. BFSP1 is myristoylated, a post-translation modification that requires caspase cleavage at D433. Bioinformatic analyses suggested that the sequences 434–452 were α-helical and amphipathic. Methods and Results: By CD spectroscopy, we show that the addition of trifluoroethanol induced a switch from an intrinsically disordered to a more α-helical conformation for the residues 434–467. Recombinantly produced BFSP1 fragments containing this amphipathic helix bind to lens lipid bilayers as determined by surface plasmon resonance (SPR). Lastly, we demonstrate by transient transfection of non-lens MCF7 cells that these same BFSP1 C-terminal sequences localise to plasma membranes and to cytoplasmic vesicles. These can be co-labelled with the vital dye, lysotracker, but other cell compartments, such as the nuclear and mitochondrial membranes, were negative. The N-terminal myristoylation of the amphipathic helix appeared not to change either the lipid affinity or membrane localisation of the BFSP1 polypeptides or fragments we assessed by SPR and transient transfection, but it did appear to enhance its helical content. Conclusions: These data support the conclusion that C-terminal sequences of human BFSP1 distal to the caspase site at G433 have independent membrane binding properties via an adjacent amphipathic helix. |
| first_indexed | 2024-03-11T02:38:50Z |
| format | Article |
| id | doaj.art-68767e82437f4ced852053678e6c9513 |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-11T02:38:50Z |
| publishDate | 2023-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-68767e82437f4ced852053678e6c95132023-11-18T09:43:01ZengMDPI AGCells2073-44092023-06-011212158010.3390/cells12121580Independent Membrane Binding Properties of the Caspase Generated Fragments of the Beaded Filament Structural Protein 1 (BFSP1) Involves an Amphipathic HelixMiguel Jarrin0Alexia A. Kalligeraki1Alice Uwineza2Chris S. Cawood3Adrian P. Brown4Edward N. Ward5Khoa Le6Stefanie Freitag-Pohl7Ehmke Pohl8Bence Kiss9Antal Tapodi10Roy A. Quinlan11Department of Biosciences, Upper Mountjoy Science Site, The University of Durham, South Road, Durham DH1 3LE, UKDepartment of Biosciences, Upper Mountjoy Science Site, The University of Durham, South Road, Durham DH1 3LE, UKDepartment of Biosciences, Upper Mountjoy Science Site, The University of Durham, South Road, Durham DH1 3LE, UKDepartment of Biosciences, Upper Mountjoy Science Site, The University of Durham, South Road, Durham DH1 3LE, UKDepartment of Biosciences, Upper Mountjoy Science Site, The University of Durham, South Road, Durham DH1 3LE, UKDepartment of Biosciences, Upper Mountjoy Science Site, The University of Durham, South Road, Durham DH1 3LE, UKBiophysical Sciences Institute, Durham University, Upper Mountjoy, South Road, Durham DH1 3LE, UKDepartment of Chemistry, Durham University, Lower Mountjoy, South Road, Durham DH1 3LE, UKBiophysical Sciences Institute, Durham University, Upper Mountjoy, South Road, Durham DH1 3LE, UKDepartment of Biochemistry and Medical Chemistry, Medical School, University of Pécs, 7624 Pécs, HungaryDepartment of Biosciences, Upper Mountjoy Science Site, The University of Durham, South Road, Durham DH1 3LE, UKDepartment of Biosciences, Upper Mountjoy Science Site, The University of Durham, South Road, Durham DH1 3LE, UKBackground: BFSP1 (beaded filament structural protein 1) is a plasma membrane, Aquaporin 0 (AQP0/MIP)-associated intermediate filament protein expressed in the eye lens. BFSP1 is myristoylated, a post-translation modification that requires caspase cleavage at D433. Bioinformatic analyses suggested that the sequences 434–452 were α-helical and amphipathic. Methods and Results: By CD spectroscopy, we show that the addition of trifluoroethanol induced a switch from an intrinsically disordered to a more α-helical conformation for the residues 434–467. Recombinantly produced BFSP1 fragments containing this amphipathic helix bind to lens lipid bilayers as determined by surface plasmon resonance (SPR). Lastly, we demonstrate by transient transfection of non-lens MCF7 cells that these same BFSP1 C-terminal sequences localise to plasma membranes and to cytoplasmic vesicles. These can be co-labelled with the vital dye, lysotracker, but other cell compartments, such as the nuclear and mitochondrial membranes, were negative. The N-terminal myristoylation of the amphipathic helix appeared not to change either the lipid affinity or membrane localisation of the BFSP1 polypeptides or fragments we assessed by SPR and transient transfection, but it did appear to enhance its helical content. Conclusions: These data support the conclusion that C-terminal sequences of human BFSP1 distal to the caspase site at G433 have independent membrane binding properties via an adjacent amphipathic helix.https://www.mdpi.com/2073-4409/12/12/1580lenscataractintermediate filamentsamphipathic helixoligomerisationintrinsically disordered domain |
| spellingShingle | Miguel Jarrin Alexia A. Kalligeraki Alice Uwineza Chris S. Cawood Adrian P. Brown Edward N. Ward Khoa Le Stefanie Freitag-Pohl Ehmke Pohl Bence Kiss Antal Tapodi Roy A. Quinlan Independent Membrane Binding Properties of the Caspase Generated Fragments of the Beaded Filament Structural Protein 1 (BFSP1) Involves an Amphipathic Helix Cells lens cataract intermediate filaments amphipathic helix oligomerisation intrinsically disordered domain |
| title | Independent Membrane Binding Properties of the Caspase Generated Fragments of the Beaded Filament Structural Protein 1 (BFSP1) Involves an Amphipathic Helix |
| title_full | Independent Membrane Binding Properties of the Caspase Generated Fragments of the Beaded Filament Structural Protein 1 (BFSP1) Involves an Amphipathic Helix |
| title_fullStr | Independent Membrane Binding Properties of the Caspase Generated Fragments of the Beaded Filament Structural Protein 1 (BFSP1) Involves an Amphipathic Helix |
| title_full_unstemmed | Independent Membrane Binding Properties of the Caspase Generated Fragments of the Beaded Filament Structural Protein 1 (BFSP1) Involves an Amphipathic Helix |
| title_short | Independent Membrane Binding Properties of the Caspase Generated Fragments of the Beaded Filament Structural Protein 1 (BFSP1) Involves an Amphipathic Helix |
| title_sort | independent membrane binding properties of the caspase generated fragments of the beaded filament structural protein 1 bfsp1 involves an amphipathic helix |
| topic | lens cataract intermediate filaments amphipathic helix oligomerisation intrinsically disordered domain |
| url | https://www.mdpi.com/2073-4409/12/12/1580 |
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