The data of heterologous expression protocol for synthesis of (15)N, (13)C-labeled SEM1(68-107) peptide fragment of homo sapiens semenogelin 1

The semenogelin 1 protein is secreted in the seminal vesicles. After ejaculation it is split into small peptide fragments using internal proteases. It was shown that the fragments SEM1(45-107), SEM1(49-107), SEM1(68-107) (SEM1(86-107) form amyloid fibrils, which increase the possibility of HIV infec...

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Autores principales: Bikmullin, Aydar, Klochkova, Evelina, Krasnovid, Filipp, Blokhin, Dmitriy
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8563649/
https://www.ncbi.nlm.nih.gov/pubmed/34754783
http://dx.doi.org/10.1016/j.mex.2021.101512
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author Bikmullin, Aydar
Klochkova, Evelina
Krasnovid, Filipp
Blokhin, Dmitriy
author_facet Bikmullin, Aydar
Klochkova, Evelina
Krasnovid, Filipp
Blokhin, Dmitriy
author_sort Bikmullin, Aydar
collection PubMed
description The semenogelin 1 protein is secreted in the seminal vesicles. After ejaculation it is split into small peptide fragments using internal proteases. It was shown that the fragments SEM1(45-107), SEM1(49-107), SEM1(68-107) (SEM1(86-107) form amyloid fibrils, which increase the possibility of HIV infection. The article presents a protocol for the synthesis and purification of a (15)N, (13)C-labeled SEM1(68–107) peptide for further structural studies by high-resolution NMR spectroscopy. The work describes cloning, expression of fusion protein GB1-SEM1(68-107) in E.coli, its purification, removal of GB1 and purification of SEM1(68-107). The purity of SEM1(68-107) samples on each purification steps was evaluated by polyacrylamide gel electrophoresis under denaturing conditions (SDS-PAGE) and tricine-SDS-PAGE. The developed protocol allows to obtain SEM1(68-107) peptide for NMR studies (using 3D experiments), instead of costly solid-phase synthesis.
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spelling pubmed-85636492021-11-08 The data of heterologous expression protocol for synthesis of (15)N, (13)C-labeled SEM1(68-107) peptide fragment of homo sapiens semenogelin 1 Bikmullin, Aydar Klochkova, Evelina Krasnovid, Filipp Blokhin, Dmitriy MethodsX Protocol Article The semenogelin 1 protein is secreted in the seminal vesicles. After ejaculation it is split into small peptide fragments using internal proteases. It was shown that the fragments SEM1(45-107), SEM1(49-107), SEM1(68-107) (SEM1(86-107) form amyloid fibrils, which increase the possibility of HIV infection. The article presents a protocol for the synthesis and purification of a (15)N, (13)C-labeled SEM1(68–107) peptide for further structural studies by high-resolution NMR spectroscopy. The work describes cloning, expression of fusion protein GB1-SEM1(68-107) in E.coli, its purification, removal of GB1 and purification of SEM1(68-107). The purity of SEM1(68-107) samples on each purification steps was evaluated by polyacrylamide gel electrophoresis under denaturing conditions (SDS-PAGE) and tricine-SDS-PAGE. The developed protocol allows to obtain SEM1(68-107) peptide for NMR studies (using 3D experiments), instead of costly solid-phase synthesis. Elsevier 2021-09-08 /pmc/articles/PMC8563649/ /pubmed/34754783 http://dx.doi.org/10.1016/j.mex.2021.101512 Text en © 2021 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol Article
Bikmullin, Aydar
Klochkova, Evelina
Krasnovid, Filipp
Blokhin, Dmitriy
The data of heterologous expression protocol for synthesis of (15)N, (13)C-labeled SEM1(68-107) peptide fragment of homo sapiens semenogelin 1
title The data of heterologous expression protocol for synthesis of (15)N, (13)C-labeled SEM1(68-107) peptide fragment of homo sapiens semenogelin 1
title_full The data of heterologous expression protocol for synthesis of (15)N, (13)C-labeled SEM1(68-107) peptide fragment of homo sapiens semenogelin 1
title_fullStr The data of heterologous expression protocol for synthesis of (15)N, (13)C-labeled SEM1(68-107) peptide fragment of homo sapiens semenogelin 1
title_full_unstemmed The data of heterologous expression protocol for synthesis of (15)N, (13)C-labeled SEM1(68-107) peptide fragment of homo sapiens semenogelin 1
title_short The data of heterologous expression protocol for synthesis of (15)N, (13)C-labeled SEM1(68-107) peptide fragment of homo sapiens semenogelin 1
title_sort data of heterologous expression protocol for synthesis of (15)n, (13)c-labeled sem1(68-107) peptide fragment of homo sapiens semenogelin 1
topic Protocol Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8563649/
https://www.ncbi.nlm.nih.gov/pubmed/34754783
http://dx.doi.org/10.1016/j.mex.2021.101512
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