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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Detalles Bibliográficos
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
Descripción
Sumario: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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