Cargando…
Effect and Mechanism of Mycobacterium tuberculosis Lipoprotein LpqH in NLRP3 Inflammasome Activation in Mouse Ana-1 Macrophage
The study is aimed at investigating the role and mechanism of LpqH of Mycobacterium tuberculosis in the activation of NLRP3 inflammasome in mouse Ana-1 macrophages. ExPASy-ProtParam, PHYRE2, ABCpred, and SYFPEITHI were used to predict and analyze the physicochemical properties, protein structure, an...
Autores principales: | , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Hindawi
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7803426/ https://www.ncbi.nlm.nih.gov/pubmed/33490276 http://dx.doi.org/10.1155/2021/8239135 |
_version_ | 1783635935740559360 |
---|---|
author | Liu, Liting Zhai, Kaixin Chen, Yue Chen, Xiaowen Wang, Guofu Wu, Lixian |
author_facet | Liu, Liting Zhai, Kaixin Chen, Yue Chen, Xiaowen Wang, Guofu Wu, Lixian |
author_sort | Liu, Liting |
collection | PubMed |
description | The study is aimed at investigating the role and mechanism of LpqH of Mycobacterium tuberculosis in the activation of NLRP3 inflammasome in mouse Ana-1 macrophages. ExPASy-ProtParam, PHYRE2, ABCpred, and SYFPEITHI were used to predict and analyze the physicochemical properties, protein structure, and B cell/T cell-associated epitopes of LpqH protein. The recombinant LpqH protein was purified, and its immunoreactivity was analyzed with western blot. The LPS-treated mouse Ana-1 macrophages were incubated with purified LpqH protein directly. The expression of NLRP3, ASC, and caspase-1 protein was detected by western blot. The secretion of IL-1β was detected by ELISA, and LDH was detected by a kit. Cell death was detected by flow cytometry. LpqH consisted of 159 amino acids and was a hydrophobic protein with stable properties. Its secondary structure contained 47% random coils, 53% β-sheets, and 3% α-helix. The tertiary structure showed a relatively loose spatial conformation. Additionally, it had 8 B cell epitopes (score > 0.8) and 10 CTL cell epitopes (score ≥ 20). The recombinant LpqH, which had strong immunoreactivity, significantly increased the levels of NLRP3, ASC, and caspase-1 p20 (P < 0.01) and promoted the secretion of IL-1β by the cells (P < 0.01). In addition, high concentration of KCl significantly inhibited the effect of LpqH on mouse Ana-1 macrophages and reduced the expression of NLRP3, ASC, and caspase-1 p20 (P < 0.01). However, there was no significant change in LDH (P > 0.05). Meanwhile, LpqH protein did not cause additional cell death (P > 0.05). LpqH protein has good immunogenicity and can activate the NLRP3 inflammasome through the potassium efflux pathway without causing cell death. |
format | Online Article Text |
id | pubmed-7803426 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Hindawi |
record_format | MEDLINE/PubMed |
spelling | pubmed-78034262021-01-22 Effect and Mechanism of Mycobacterium tuberculosis Lipoprotein LpqH in NLRP3 Inflammasome Activation in Mouse Ana-1 Macrophage Liu, Liting Zhai, Kaixin Chen, Yue Chen, Xiaowen Wang, Guofu Wu, Lixian Biomed Res Int Research Article The study is aimed at investigating the role and mechanism of LpqH of Mycobacterium tuberculosis in the activation of NLRP3 inflammasome in mouse Ana-1 macrophages. ExPASy-ProtParam, PHYRE2, ABCpred, and SYFPEITHI were used to predict and analyze the physicochemical properties, protein structure, and B cell/T cell-associated epitopes of LpqH protein. The recombinant LpqH protein was purified, and its immunoreactivity was analyzed with western blot. The LPS-treated mouse Ana-1 macrophages were incubated with purified LpqH protein directly. The expression of NLRP3, ASC, and caspase-1 protein was detected by western blot. The secretion of IL-1β was detected by ELISA, and LDH was detected by a kit. Cell death was detected by flow cytometry. LpqH consisted of 159 amino acids and was a hydrophobic protein with stable properties. Its secondary structure contained 47% random coils, 53% β-sheets, and 3% α-helix. The tertiary structure showed a relatively loose spatial conformation. Additionally, it had 8 B cell epitopes (score > 0.8) and 10 CTL cell epitopes (score ≥ 20). The recombinant LpqH, which had strong immunoreactivity, significantly increased the levels of NLRP3, ASC, and caspase-1 p20 (P < 0.01) and promoted the secretion of IL-1β by the cells (P < 0.01). In addition, high concentration of KCl significantly inhibited the effect of LpqH on mouse Ana-1 macrophages and reduced the expression of NLRP3, ASC, and caspase-1 p20 (P < 0.01). However, there was no significant change in LDH (P > 0.05). Meanwhile, LpqH protein did not cause additional cell death (P > 0.05). LpqH protein has good immunogenicity and can activate the NLRP3 inflammasome through the potassium efflux pathway without causing cell death. Hindawi 2021-01-04 /pmc/articles/PMC7803426/ /pubmed/33490276 http://dx.doi.org/10.1155/2021/8239135 Text en Copyright © 2021 Liting Liu et al. https://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Article Liu, Liting Zhai, Kaixin Chen, Yue Chen, Xiaowen Wang, Guofu Wu, Lixian Effect and Mechanism of Mycobacterium tuberculosis Lipoprotein LpqH in NLRP3 Inflammasome Activation in Mouse Ana-1 Macrophage |
title | Effect and Mechanism of Mycobacterium tuberculosis Lipoprotein LpqH in NLRP3 Inflammasome Activation in Mouse Ana-1 Macrophage |
title_full | Effect and Mechanism of Mycobacterium tuberculosis Lipoprotein LpqH in NLRP3 Inflammasome Activation in Mouse Ana-1 Macrophage |
title_fullStr | Effect and Mechanism of Mycobacterium tuberculosis Lipoprotein LpqH in NLRP3 Inflammasome Activation in Mouse Ana-1 Macrophage |
title_full_unstemmed | Effect and Mechanism of Mycobacterium tuberculosis Lipoprotein LpqH in NLRP3 Inflammasome Activation in Mouse Ana-1 Macrophage |
title_short | Effect and Mechanism of Mycobacterium tuberculosis Lipoprotein LpqH in NLRP3 Inflammasome Activation in Mouse Ana-1 Macrophage |
title_sort | effect and mechanism of mycobacterium tuberculosis lipoprotein lpqh in nlrp3 inflammasome activation in mouse ana-1 macrophage |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7803426/ https://www.ncbi.nlm.nih.gov/pubmed/33490276 http://dx.doi.org/10.1155/2021/8239135 |
work_keys_str_mv | AT liuliting effectandmechanismofmycobacteriumtuberculosislipoproteinlpqhinnlrp3inflammasomeactivationinmouseana1macrophage AT zhaikaixin effectandmechanismofmycobacteriumtuberculosislipoproteinlpqhinnlrp3inflammasomeactivationinmouseana1macrophage AT chenyue effectandmechanismofmycobacteriumtuberculosislipoproteinlpqhinnlrp3inflammasomeactivationinmouseana1macrophage AT chenxiaowen effectandmechanismofmycobacteriumtuberculosislipoproteinlpqhinnlrp3inflammasomeactivationinmouseana1macrophage AT wangguofu effectandmechanismofmycobacteriumtuberculosislipoproteinlpqhinnlrp3inflammasomeactivationinmouseana1macrophage AT wulixian effectandmechanismofmycobacteriumtuberculosislipoproteinlpqhinnlrp3inflammasomeactivationinmouseana1macrophage |