The hydrophobic C‐terminal sequence of transthyretin affects its catalytic kinetics towards amidated neuropeptide Y
Transthyretin (TTR) is a transporter for thyroid hormone and retinol binding protein that has recently been reported to have proteolytic activity against certain substrates, including amidated neuropeptide Y (NPY). However, the proteolytic activity of TTR towards NPY is not fully understood. Here, w...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Wiley
2019-04-01
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| Series: | FEBS Open Bio |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/2211-5463.12604 |
| _version_ | 1797821931523670016 |
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| author | Sukanya Tangthavewattana Ladda Leelawatwattana Porntip Prapunpoj |
| author_facet | Sukanya Tangthavewattana Ladda Leelawatwattana Porntip Prapunpoj |
| author_sort | Sukanya Tangthavewattana |
| collection | DOAJ |
| description | Transthyretin (TTR) is a transporter for thyroid hormone and retinol binding protein that has recently been reported to have proteolytic activity against certain substrates, including amidated neuropeptide Y (NPY). However, the proteolytic activity of TTR towards NPY is not fully understood. Here, we used fluorescence‐based assays to determine the catalytic kinetics of human TTR towards human amidated NPY. The Michaelis constant (KM) and catalytic efficiency (kcat/KM) of TTR proteolysis were 15.88 ± 0.44 μm and 687 081 ± 35 692 m−1·s−1, respectively. In addition, we demonstrated an effect of the C‐terminal sequence of TTR. When the C‐terminal sequence of TTR was made more hydrophobic, the KM and kcat/KM changed to 12.87 ± 0.22 μm and 983 755 ± 18 704 m−1·s−1, respectively. Our results may be useful for the development of TTR as a therapeutic agent with low risk of the undesirable symptoms that develop from amidated NPY, and for further improvement of the kcat/KM of TTR. |
| first_indexed | 2024-03-13T09:59:58Z |
| format | Article |
| id | doaj.art-9ed91b4c05f74fac99e303b512eff0e5 |
| institution | Directory Open Access Journal |
| issn | 2211-5463 |
| language | English |
| last_indexed | 2024-03-13T09:59:58Z |
| publishDate | 2019-04-01 |
| publisher | Wiley |
| record_format | Article |
| series | FEBS Open Bio |
| spelling | doaj.art-9ed91b4c05f74fac99e303b512eff0e52023-05-23T06:46:21ZengWileyFEBS Open Bio2211-54632019-04-019459460410.1002/2211-5463.12604The hydrophobic C‐terminal sequence of transthyretin affects its catalytic kinetics towards amidated neuropeptide YSukanya Tangthavewattana0Ladda Leelawatwattana1Porntip Prapunpoj2Department of Biochemistry Faculty of Science Prince of Songkla University Hat Yai ThailandDepartment of Biochemistry Faculty of Science Prince of Songkla University Hat Yai ThailandDepartment of Biochemistry Faculty of Science Prince of Songkla University Hat Yai ThailandTransthyretin (TTR) is a transporter for thyroid hormone and retinol binding protein that has recently been reported to have proteolytic activity against certain substrates, including amidated neuropeptide Y (NPY). However, the proteolytic activity of TTR towards NPY is not fully understood. Here, we used fluorescence‐based assays to determine the catalytic kinetics of human TTR towards human amidated NPY. The Michaelis constant (KM) and catalytic efficiency (kcat/KM) of TTR proteolysis were 15.88 ± 0.44 μm and 687 081 ± 35 692 m−1·s−1, respectively. In addition, we demonstrated an effect of the C‐terminal sequence of TTR. When the C‐terminal sequence of TTR was made more hydrophobic, the KM and kcat/KM changed to 12.87 ± 0.22 μm and 983 755 ± 18 704 m−1·s−1, respectively. Our results may be useful for the development of TTR as a therapeutic agent with low risk of the undesirable symptoms that develop from amidated NPY, and for further improvement of the kcat/KM of TTR.https://doi.org/10.1002/2211-5463.12604amidated neuropeptide YC‐terminal sequencefluorescence‐based assaykineticsproteolytic activitytransthyretin |
| spellingShingle | Sukanya Tangthavewattana Ladda Leelawatwattana Porntip Prapunpoj The hydrophobic C‐terminal sequence of transthyretin affects its catalytic kinetics towards amidated neuropeptide Y FEBS Open Bio amidated neuropeptide Y C‐terminal sequence fluorescence‐based assay kinetics proteolytic activity transthyretin |
| title | The hydrophobic C‐terminal sequence of transthyretin affects its catalytic kinetics towards amidated neuropeptide Y |
| title_full | The hydrophobic C‐terminal sequence of transthyretin affects its catalytic kinetics towards amidated neuropeptide Y |
| title_fullStr | The hydrophobic C‐terminal sequence of transthyretin affects its catalytic kinetics towards amidated neuropeptide Y |
| title_full_unstemmed | The hydrophobic C‐terminal sequence of transthyretin affects its catalytic kinetics towards amidated neuropeptide Y |
| title_short | The hydrophobic C‐terminal sequence of transthyretin affects its catalytic kinetics towards amidated neuropeptide Y |
| title_sort | hydrophobic c terminal sequence of transthyretin affects its catalytic kinetics towards amidated neuropeptide y |
| topic | amidated neuropeptide Y C‐terminal sequence fluorescence‐based assay kinetics proteolytic activity transthyretin |
| url | https://doi.org/10.1002/2211-5463.12604 |
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