Selective recognition of phosphatidylinositol phosphate receptors by C‑terminal tail of mitotic kinesin-like protein 2 (MKlp2)
The mitotic kinesin-like protein 2 (MKlp2) plays a key role in the proper completion of cytokinetic abscission. Specifically, the C-terminal tail of MKlp2 (CTM peptides) offers a stable tethering on the plasma membrane and microtubule cytoskeleton in the midbody during abscission. However, little is...
| Main Authors: | , , , , |
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| Other Authors: | |
| Format: | Journal Article |
| Language: | English |
| Published: |
2022
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| Subjects: | |
| Online Access: | https://hdl.handle.net/10356/162153 |
| _version_ | 1811686756615979008 |
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| author | Tae, Hyunhyuk Park, Soohyun Kim, Seong-Oh Yorulmaz, Saziye Cho, Nam-Joon |
| author2 | School of Materials Science and Engineering |
| author_facet | School of Materials Science and Engineering Tae, Hyunhyuk Park, Soohyun Kim, Seong-Oh Yorulmaz, Saziye Cho, Nam-Joon |
| author_sort | Tae, Hyunhyuk |
| collection | NTU |
| description | The mitotic kinesin-like protein 2 (MKlp2) plays a key role in the proper completion of cytokinetic abscission. Specifically, the C-terminal tail of MKlp2 (CTM peptides) offers a stable tethering on the plasma membrane and microtubule cytoskeleton in the midbody during abscission. However, little is known about the underlying mechanism of how the CTM peptides bind to the plasma membrane of the intercellular bridge. Herein, we identify the specific molecular interaction between the CTM peptides and phosphatidylinositol phosphate (PIP) receptors using quartz crystal microbalance-dissipation and atomic force microscopy force spectroscopic measurements. To systematically examine the effects of amino acids, we designed a series of synthetic 33-mer peptides derived from the wild-type (CTM1). First, we evaluated the peptide binding amount caused by electrostatic interactions based on 100% zwitterionic and 30% negatively charged model membranes, whereby the nonspecific attractions were nearly proportional to the net charge of peptides. Upon incubating with PIP-containing model membranes, the wild-type CTM1 and its truncated mutation showed significant PI(3)P-specific binding, which was evidenced by a 15-fold higher binding mass and 6-fold stronger adhesion force compared to other negatively charged membranes. The extent of the specific binding was predominantly dependent on the existence of S21, whereby substitution or deletion of S21 significantly hindered the binding affinity. Taken together, our findings based on a correlative measurement platform enabled the quantification of the nonelectrostatic, selective binding interactions of the C-terminal of MKlp2 to certain PIP receptors and contributed to understanding the molecular mechanisms on complete cytokinetic abscission in cells. |
| first_indexed | 2024-10-01T05:05:29Z |
| format | Journal Article |
| id | ntu-10356/162153 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T05:05:29Z |
| publishDate | 2022 |
| record_format | dspace |
| spelling | ntu-10356/1621532022-10-06T02:31:31Z Selective recognition of phosphatidylinositol phosphate receptors by C‑terminal tail of mitotic kinesin-like protein 2 (MKlp2) Tae, Hyunhyuk Park, Soohyun Kim, Seong-Oh Yorulmaz, Saziye Cho, Nam-Joon School of Materials Science and Engineering China-Singapore International Joint Research Institute (CSIJRI) Engineering::Maritime studies Binding Energy Cell Membranes The mitotic kinesin-like protein 2 (MKlp2) plays a key role in the proper completion of cytokinetic abscission. Specifically, the C-terminal tail of MKlp2 (CTM peptides) offers a stable tethering on the plasma membrane and microtubule cytoskeleton in the midbody during abscission. However, little is known about the underlying mechanism of how the CTM peptides bind to the plasma membrane of the intercellular bridge. Herein, we identify the specific molecular interaction between the CTM peptides and phosphatidylinositol phosphate (PIP) receptors using quartz crystal microbalance-dissipation and atomic force microscopy force spectroscopic measurements. To systematically examine the effects of amino acids, we designed a series of synthetic 33-mer peptides derived from the wild-type (CTM1). First, we evaluated the peptide binding amount caused by electrostatic interactions based on 100% zwitterionic and 30% negatively charged model membranes, whereby the nonspecific attractions were nearly proportional to the net charge of peptides. Upon incubating with PIP-containing model membranes, the wild-type CTM1 and its truncated mutation showed significant PI(3)P-specific binding, which was evidenced by a 15-fold higher binding mass and 6-fold stronger adhesion force compared to other negatively charged membranes. The extent of the specific binding was predominantly dependent on the existence of S21, whereby substitution or deletion of S21 significantly hindered the binding affinity. Taken together, our findings based on a correlative measurement platform enabled the quantification of the nonelectrostatic, selective binding interactions of the C-terminal of MKlp2 to certain PIP receptors and contributed to understanding the molecular mechanisms on complete cytokinetic abscission in cells. Agency for Science, Technology and Research (A*STAR) Ministry of Education (MOE) This work was supported by the Ministry of Education (MOE) in Singapore under grant AcRF TIER1-2020-T1-002-032 (RG111/20). This work also supported by the ChinaSingapore International Joint Research Institute (CSIJRI). Figures 2 A, 3 A, and 4 A were created with BioRender.com. H.T. is supported by an SINGA graduate scholarship from the A*STAR Graduate Academy, Singapore. 2022-10-06T02:31:31Z 2022-10-06T02:31:31Z 2022 Journal Article Tae, H., Park, S., Kim, S., Yorulmaz, S. & Cho, N. (2022). Selective recognition of phosphatidylinositol phosphate receptors by C‑terminal tail of mitotic kinesin-like protein 2 (MKlp2). Journal of Physical Chemistry B, 126(12), 2345-2352. https://dx.doi.org/10.1021/acs.jpcb.1c10534 1520-6106 https://hdl.handle.net/10356/162153 10.1021/acs.jpcb.1c10534 35316051 2-s2.0-85127677989 12 126 2345 2352 en Journal of Physical Chemistry B © 2022 American Chemical Society. All rights reserved. |
| spellingShingle | Engineering::Maritime studies Binding Energy Cell Membranes Tae, Hyunhyuk Park, Soohyun Kim, Seong-Oh Yorulmaz, Saziye Cho, Nam-Joon Selective recognition of phosphatidylinositol phosphate receptors by C‑terminal tail of mitotic kinesin-like protein 2 (MKlp2) |
| title | Selective recognition of phosphatidylinositol phosphate receptors by C‑terminal tail of mitotic kinesin-like protein 2 (MKlp2) |
| title_full | Selective recognition of phosphatidylinositol phosphate receptors by C‑terminal tail of mitotic kinesin-like protein 2 (MKlp2) |
| title_fullStr | Selective recognition of phosphatidylinositol phosphate receptors by C‑terminal tail of mitotic kinesin-like protein 2 (MKlp2) |
| title_full_unstemmed | Selective recognition of phosphatidylinositol phosphate receptors by C‑terminal tail of mitotic kinesin-like protein 2 (MKlp2) |
| title_short | Selective recognition of phosphatidylinositol phosphate receptors by C‑terminal tail of mitotic kinesin-like protein 2 (MKlp2) |
| title_sort | selective recognition of phosphatidylinositol phosphate receptors by c terminal tail of mitotic kinesin like protein 2 mklp2 |
| topic | Engineering::Maritime studies Binding Energy Cell Membranes |
| url | https://hdl.handle.net/10356/162153 |
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