Cloning, Expression, and Characterization of Der f 7, an Allergen of<i>Dermatophagoides farinae</i>From China

Cui, Yubao; Cai, Hua; Zhou, Ying; Gao, Cuixiang; Shi, Wenqi; Yu, Mengying; Li, Li

doi:10.1093/jmedent/47.5.868

Cited by 18 publications

(6 citation statements)

References 14 publications

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“…After confirmation by restriction enzyme digests and DNA sequencing, the expression plasmid was transformed into the BL21 E. coli strain. A high expression strain was selected and induced and the expressed recombinant protein was purified using MBP (maltose-binding proteins) affinity14,15 More information is shown in Figure S1.…”

Section: Methodsmentioning

confidence: 99%

Arsenic trioxide inhibits viability of pancreatic cancer stem cells in culture and in a xenograft model via binding to SHH-Gli

Han¹,

Sang²,

Chang³

et al. 2013

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ObjectiveOverexpression of the sonic hedgehog (SHH) signaling pathway is an essential characteristic of pancreatic cancer stem cells (PCSCs) and arsenic trioxide (ATO) is described as a SHH inhibitor. This study evaluates whether ATO has the potential to inhibit viability of PCSCs via binding to SHH-Gli proteins.MethodsCell counting kit-8 and flow cytometry were used for analyzing apoptosis in cells in vitro. The animal model was an athymic nude mouse model bearing subcutaneous xenografts of SW1990 pancreatic cancer cells. The terminal deoxynucleotidyl transferase dUTP nick end labeling assay and immunohistochemistry were used for tumor tissue analysis. The interaction between Gli1 and ATO was examined by a confocal system and an ultraviolet absorption spectrum assay.ResultsATO induced apoptosis in pancreatic cancer cells, especially CD24+CD44+ cells in vitro. Combination treatment of ATO and low dose gemcitabine inhibited tumor growth by 60.9% (P = 0.004), and decreased the expression of CD24, CD44, and aldehyde dehydrogenase 1 family, member A1 significantly in vivo. ATO changed the structure of the recombinant Gli1 zinc finger peptides in a cell-free condition and the binding action of ATO to recombinant Gli1 was observed in cultured pancreatic cancer cells.ConclusionATO may have the potential to inhibit viability of PCSCs via binding to SHH-Gli proteins in vitro and in vivo.

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Section: Methodsmentioning

confidence: 99%

Arsenic trioxide inhibits viability of pancreatic cancer stem cells in culture and in a xenograft model via binding to SHH-Gli

Han¹,

Sang²,

Chang³

et al. 2013

OTT

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“…Monoclonal antibodies (mAb) produced against Der p 7 and Der f 7 have cross-reacted with the group 7 allergens of both species and blocked IgE binding to these allergens [6], [7], suggesting that Der f 7 and Der p 7 may share similar IgE epitopes. An isoform of Der f 7 has been cloned, expressed and the secondary structure characterized [8], but the detailed three-dimensional structure of Der f 7 is not available. A three-dimensional model of Der f 7 was generated using homology modeling, based on the crystal structure of Der p 7 and used for mAb binding studies [9].…”

Section: Introductionmentioning

confidence: 99%

Crystal Structure of Der f 7, a Dust Mite Allergen from Dermatophagoides farinae

et al. 2012

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BackgroundDer f 7 is the group 7 allergen from the dust mite Dermatophagoides farinae, homologous to the major allergen Der p 7 from D. pteronyssinus. Monoclonal antibody that bind to residues Leu48 and Phe50 was found to inhibit IgE binding to residue Asp159, which is important for the cross-reactivity between Der f 7 and Der p 7.Methodology/Principal FindingsHere, we report the crystal structure of Der f 7 that shows an elongated and curved molecule consisting of two anti-parallel β-sheets – one 4-stranded and the other 5-stranded – that wrap around a long C-terminal helix. The overall fold of Der f 7 is similar to Der p 7 but key difference was found in the β1–β2 loop region. In Der f 7, Leu48 and Phe50 are in close proximity to Asp159, explaining why monoclonal antibody binding to Leu48 and Phe50 can inhibit IgE binding to Asp159. Both Der f 7 and Der p 7 bind weakly to polymyxin B via a similar binding site that is formed by the N-terminal helix, the 4-stranded β-sheet and the C-terminal helix. The thermal stability of Der f 7 is significantly lower than that of Der p 7, and the stabilities of both allergens are highly depend on pH.Conclusion/SignificanceDer f 7 is homologous to Der p 7 in terms of the amino acid sequence and overall 3D structure but with significant differences in the region proximal to the IgE epitope and in thermal stability. The crystal structure of Der f 7 provides a basis for studying the function and allergenicity of this group of allergens.

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“…In biochemistry and structural biology, secondary protein structure is the general three-dimensional form of local segments of proteins, defined by patterns of hydrogen bonds between backbone amide and carboxyl groups (20). By the GOR IV program, the secondary structure of the mature protein is composed of alpha helices (24.86%), extended strands (27.01%), and random coils (48.13%).…”

Section: Discussionmentioning

confidence: 99%

Cloning of the surface layer gene sllB from Bacillus sphaericus ATCC 14577 and its heterologous expression and purification

Cui

Zhou

Liu³

et al. 2012

International Journal of Molecular Medicine

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A cDNA fragment encoding the S-layer protein SllB cloned from Bacillus sphaericus ATCC 14577 was expressed on the surface of E. coli BL21 (DE3) cells and confirmed by the square lattice structure at the nanoscale level. The amplified gene fragment designed with PCR primers from a specified reference sequence (GenBank accession no. AJ849550) showed a high degree of sequence identity with the known sequences for S-layer protein. The best alignment scores were seen in B. sphaericus strains JG-A12 and NCTC9602, which code for a pre-form protein with a predicted cleavage site located between the two alanine residues 31 and 32. After this signal peptide sequence was removed, the mature protein had a molecular mass of 116.2613 kDa and a theoretical pI of 5.40. Further bioinformatic analysis revealed three S-layer homology (SLH) domains in the N-terminus of the mature protein, positioned at the 1–61, 63–128 and 137–197 residues. The mature S-layer protein was composed of alpha helices (24.86%), extended strands (27.01%), and rich random coils (48.13%). Bioinformatics-driven characterization of SllB may provide scientific evidence for further application of this gene in the fields of nanobiotechnology and biomimetics in the future.

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Cloning, Expression, and Characterization of Der f 7, an Allergen ofDermatophagoides farinaeFrom China

Cited by 18 publications

References 14 publications

Arsenic trioxide inhibits viability of pancreatic cancer stem cells in culture and in a xenograft model via binding to SHH-Gli

Arsenic trioxide inhibits viability of pancreatic cancer stem cells in culture and in a xenograft model via binding to SHH-Gli

Crystal Structure of Der f 7, a Dust Mite Allergen from Dermatophagoides farinae

Cloning of the surface layer gene sllB from Bacillus sphaericus ATCC 14577 and its heterologous expression and purification

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