AQUACULTURE RESEARCH
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2016
·Vol. 48
·期 5
·2278-2290
·
IF 1.606
Jing Qian,Chunhua Ren,Ting Chen,Jianjun Xia,Zonghe Yu,Yan Gao,Chaoqun Hu
The broad-spectrum protease inhibitor alpha-2-macroglobulin (α 2 M) is widely found in vertebrates and invertebrates. However, no research regarding α 2 M has been described in sea cucumbers to date. In this study, the first sea cucumber α 2 M named Stm α 2 M was identified from Stichopus monotuberculatus , a tropical species with high edible and economic values in Southern China. The Stm α 2 M cDNA is 5276 bp in length, containing a 5′-untranslated region (UTR) of 279 bp, a 3′-UTR of 344 bp and an open reading frame of 4653 bp that encodes a protein of 1550 amino acids. The Stmα 2 M possesses all the three characteristics of known α 2 M proteins, including the bait region domain, thioester domain and receptor-binding domain. Higher levels of mRNA expression were noticed in intestine, respiratory tree, coelomocytes and tentacle of S. monotuberculatus . Transcriptional expression of Stm α 2 M showed the strongest response to lipopolysaccharides (LPS, 8.19-fold up-regulation) after 6 h post-challenge in coelomocytes. In addition, the α 2 M activity was detected in S. monotuberculatus and protein activity of α 2 M increased remarkably at 6 h after LPS stimulation. As a whole, this study indicated that Stmα 2 M is an important immune-related molecule, and the study on functions of Stmα 2 M may provide some new and valuable ideas for preventing and controlling diseases of cultured sea cucumbers.
研究领域:
免疫学
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分子生物学
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海洋生物学
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DOI
高影响因子
JOURNAL OF VIROLOGY
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2023
·Vol. 97
·期 5
·
IF 5.4
Renrong You,Kangchengyin Liu,Min Huang,Lihua Tang,Xuehui Zhang,Yahui Huang,Jing Zhao,Ye Zhao,Lilin Ye,Guozhong Zhang
Infectious bronchitis virus (IBV) infections are initiated by the transmembrane spike (S) glycoprotein, which binds to host factors and fuses the viral and cell membranes. The N-terminal domain of the S1 subunit of IBV S protein binds to sialic acids, but the precise location of the sialic acid binding domain (SABD) and the role of the SABD in IBV-infected chickens remain unclear. Here, we identify the S1 N-terminal amino acid (aa) residues 19 to 227 (209 aa total) of IBV strains SD (GI-19) and GD (GI-7), and the corresponding region of M41 (GI-1), as the minimal SABD using truncated protein histochemistry and neuraminidase assays. Both α-2,3- and α-2,6-linked sialic acids on the surfaces of CEK cells can be used as attachment receptors by IBV, leading to increased infection efficiency. However, 9-O acetylation of the sialic acid glycerol side chain inhibits IBV S1 and SABD protein binding. We further constructed recombinant strains in which the S1 gene or the SABD in the GD and SD genomes were replaced with the corresponding region from M41 by reverse genetics. Infecting chickens with these viruses revealed that the virulence and nephrotropism of rSDM41-S1, rSDM41-206, rGDM41-S1, and rGDM41-206 strains were decreased to various degrees compared to their parental strains. A positive sera cross-neutralization test showed that the serotypes were changed for the recombinant viruses. Our results provide insight into IBV infection of host cells that may aid vaccine design.IMPORTANCE To date, only α-2,3-linked sialic acid has been identified as a potential host binding receptor for IBV. Here, we show the minimum region constituting the sialic acid binding domain (SABD) and the binding characteristics of the S1 subunit of spike (S) protein of IBV strains SD (GI-19), GD (GI-7), and M41 (GI-1) to various sialic acids. The 9-O acetylation modification partially inhibits IBV from binding to sialic acid, while the virus can also bind to sialic acid molecules linked to host cells through an α-2,6 linkage, serving as another receptor determinant. Substitution of the putative SABD from strain M41 into strains SD and GD resulted in reduced virulence, nephrotropism, and a serotype switch. These findings suggest that sialic acid binding has diversified during the evolution of γ-coronaviruses, impacting the biological properties of IBV strains. Our results offer insight into the mechanisms by which IBV invades host cells.
研究领域:
病毒学
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分子生物学
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免疫学
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动物健康
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DOI
PubMed
高影响因子
GENE
·
2016
·
IF 2.319
Jing Qian,Chunhua Ren,Jianjun Xia,Ting Chen,Zonghe Yu,Chaoqun Hu
The non-specific protease inhibitor alpha-2-macroglobulin (A 2 M) is a key macromolecular glycoprotein that involved in host immune defense against pathogens in vertebrates and invertebrates. However, no research regarding A 2 M has been developed in echinoderms to date. In this study, the full-length cDNA of A 2 M was cloned from the sea cucumber ( Holothuria atra ), which is a tropical species widely distributed along the coasts of the South China Sea and designated HaA 2 M . HaA 2 M possesses all three conserved functional domains of known A 2 M proteins, including the bait region domain, thioester domain and receptor-binding domain. Compared to fish and shrimp A 2 Ms, the histidine residue from the catalytical regions is well conserved in HaA 2 M. HaA 2 M mRNA was predominantly expressed in coelomocytes and, to a lesser extent, in the body wall, intestine and respiratory tree. A 2 M activity was detected in the coelomic fluids of H. atra . The mRNA expression and activity levels were investigated in the major immune tissues and coelomic fluids of H. atra after challenge with inactivated Vibrio alginolyticus or polyriboinosinic polyribocytidylic acid [Poly (I: C)]. RNA interference (RNAi)-mediated knockdown of HaA 2 M resulted in a significant reduction of HaA 2 M gene transcript level (86%). RNAi-mediated silencing of HaA 2 M gene significantly decreased the A 2 M activity (38%) and increased the number of viable bacteria (2.8-fold) in the coelomic fluids of H. atra infected by V. alginolyticus . Our study, as a whole, supplied the evidences for HaA 2 M as an immune-relevant molecule and it might have multiple functions in the innate immune system of H. atra .
研究领域:
免疫学
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分子生物学
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海洋生物学
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DOI
PubMed
高影响因子
JOURNAL OF THE WORLD AQUACULTURE SOCIETY
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2016
·Vol. 48
·期 2
·360-373
·
IF 0.665
Jing Qian,Chunhua Ren,Jianjun Xia,Zonghe Yu,Chaoqun Hu
Alpha-2-macroglobulin (A 2 M) is essential for hosts defending against pathogen infections. In this study, two distinct A 2 Ms were identified from sea cucumbers Stichopus monotuberculatus and Holothuria atra , which belong to different genera in the Phylum Echinodermata, and named StmA 2 M and HaA 2 M, respectively. Structural analysis of StmA 2 M and HaA 2 M revealed that all contain the bait region domain, thioester domain, and receptor-binding domain. Phylogenetic analysis indicated that StmA 2 M and HaA 2 M all belong to the echinoderm A 2 Ms. The cleavage site RXXR and the catalytic histidine residue for complement component C3 were found in StmA 2 M and HaA 2 M. Compared with human A 2 M, the three-dimensional structures of StmA 2 M and HaA 2 M were more similar to that of common carp A 2 M. Both StmA 2 M and HaA 2 M showed the strongest response at transcriptional levels to lipopolysaccharide (LPS) at 12 h and polyriboinosinic polyribocytidylic acid [Poly (I: C)] at 24 h, respectively. Activity of StmA 2 M showed the strongest response to Poly (I: C) at 6 h, while the strongest change of HaA 2 M activity was induced by LPS at 24 h. The differential mRNA expression and enzyme activity levels suggested that A 2 M in sea cucumbers might be involved in different immune responses to LPS or Poly (I: C) challenge.
研究领域:
海洋生物学
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免疫学
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分子生物学
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基因组学
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DOI
高影响因子
MICROCHIMICA ACTA
·
2023
·Vol. 190
·期 10
·1-11
·
IF 5.7
Tian Jiangman,Zhu Qihao,Huang Xiaojing,Li Yuan
A novel sandwich-type “on-off” electrochemiluminescence (ECL) biosensor for the determination of α(2,3)-sial-Gs was designed. Specifically, amino-functionalized porous silica nanoparticles (HPSNs-NH 2 ) were first prepared and then decorated with gold nanoparticles (Au NPs) to form HPSNs-NH 2 @Au NP nanocomposite, which exhibited a strong ability to enhance ECL intensity with K 2 S 2 O 8 as co-reactant (signal-on) and could immobilize the target-specific binding molecules of maackia amurensis lectin (MAL). Additionally, AuPdPt trimetallic nanoparticles were prepared to serve as a quenched ECL signal indicator (signal-off) with the ability of capturing the target non-specific binding molecules of 3-aminophenylboronic acid (APBA) to form a signal label. The sandwich-type ECL biosensor was constructed based on the structure of MAL-α(2,3)-sial-Gs-APBA and achieved a determination toward α(2,3)-sial-Gs with a wide linear range from 1 fg mL −1 to 10 ng mL −1 and a low detection limit of 0.5 fg mL −1 . Furthermore, the proposed ECL biosensor showed satisfactory selectivity, stability, and reproducibility for α(2,3)-sial-Gs determination. Graphical Abstract
研究领域:
分析化学
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生物医学工程
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纳米科技
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DOI
PubMed
高影响因子
NEOPLASIA
·
2026
·
IF 4.8
Hongming Teng,Yuxin Guo,Xinran Chen,Anlian Fan,Chengfei Zhang,Ting Zhang,Yuanyuan Luo,Lin Huang
Colorectal cancer (CRC) progression could be fueled by the activation of the unfolded protein response (UPR) triggered by endoplasmic reticulum (ER) stress. The proline-rich Akt1 substrate of 40 kDa (PRAS40) is implicated in cancer progression, but its role in the UPR remains unclear. Herein, we demonstrate that PRAS40 promotes the inositol-requiring enzyme 1α (IRE1α)-X-box binding protein 1 (XBP1) axis-dependent UPR in driving CRC progression. Mechanistically, PRAS40 interacts with ER chaperone glucose-regulated protein 78 (GRP78) and enhances its N-glycosylation. Moreover, PRAS40 improves the interaction between GRP78 and ST6 β-galactoside α-2, 6-sialyltransferase 1 (ST6Gal1), leading to increased α-2, 6-sialylation of GRP78 and the UPR triggered by ER stress. Furthermore, we identified the natural compound β-sitosterol as a novel ST6Gal1 inhibitor, which attenuated PRAS40-triggered tumor growth. Collectively, these findings unveil a PRAS40-ST6Gal1-GRP78 axis that drives CRC progression through activating the IRE1α-XBP-1-mediated UPR and nominate ST6Gal1 as a promising therapeutic target.
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DOI
高影响因子
ANALYTICAL CHEMISTRY
·
2021
·
IF 6.986
Mengxia Cheng,Hong Shu,Ye Peng,Xiaoxiao Feng,Guoquan Yan,Lei Zhang,Jun Yao,Huimin Bao,Haojie Lu
Sialylated N-glycan isomers with α-2,3 and α-2,6 linkages play crucial and distinctive roles in diverse physiological and pathological processes. Changes of α-2,3-linked sialic acids in sialylated N-glycans are especially important in monitoring the initiation and progression of diseases. However, the specific analysis of α-2,3-sialylated N-glycan linkage isomers remains challenging due to their extremely low abundance and technical limitations in separation and detection. Herein, we designed an integrated strategy that combines linkage-specific derivatization and a charge-sensitive separation method based on microfluidic chip capillary electrophoresis–mass spectrometry (microchip CE–MS) for specific analysis of α-2,3-sialylated N-glycan linkage isomers for the first time. The α-2,6- and α-2,3-sialic acids were selectively labeled with methylamine (MA) and N,N-dimethylethylenediamine (DMEN), respectively, which selectively makes α-2,3-sialylated N-glycans positively charged and realizes online purification, concentration, and discrimination of α-2,3-sialylated N-glycans from other N-glycans in microchip CE–MS. This new approach was demonstrated with standard multisialylated N-glycans, and it was found that only the α-2,3-sialylated N-glycans migrated and were detected in order according to the number of α-2,3-sialic acids. Finally, this strategy was successfully applied in highly sensitive profiling and reproducible quantitation of the serum α-2,3-sialylated N-glycome from ovarian cancer (OC) patients, where 7 of 33 detected α-2,3-sialylated N-glycans significantly changed in the OC group compared with healthy controls.
研究领域:
分析化学
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生物化学
·
医学
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产品关联
DOI
PubMed
高影响因子
EXPERIMENTAL CELL RESEARCH
·
2024
·
IF 3.3
Jianfeng Ye,Cheng Qiu,Lexi Zhang
Renal ischemia-reperfusion (I/R) injury leads mainly to acute kidney injury. Leucine-rich alpha-2-glycoprotein 1 (LRG) is upregulated in kidney tissues of mice after renal I/R injury. However, its role in renal I/R injury has not been fully elucidated. A mouse model of renal I/R injury was constructed by unilateral renal pedicle clamping and reperfusion. Mice undergoing I/R procedures exhibited renal function impairment and increased LRG protein expression compared with mice receiving sham operations. Tail vein injection with lentivirus carrying shLRG decreased renal I/R injury-induced increase in caspase-3 activity, IL-1β and IL-18 concentrations, and ROS production. Furthermore, shRNA-mediated LRG knockdown in HK-2 cells protected against H/R-induced cell damage. LRG could upregulate the expression of NADPH oxidase 4 (NOX4). We also determined the increased NOX4 expression in kidney tissues of renal I/R-operated mice and H/R-treated HK-2 cells. NOX4 overexpression reversed the inhibitory role of LRG knockdown in HK-2 cell damage caused by H/R. Collectively, our findings demonstrate that LRG knockdown decreases the NOX4 expression, thereby alleviating renal I/R injury by inhibiting cell apoptosis, inflammation, and oxidative stress.
研究领域:
肾病学
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分子生物学
·
细胞生物学
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产品关联
DOI
PubMed
高影响因子
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