Construction of multiple recombinant SLA-I proteins by linking heavy chains and light chains in vitro and analyzing their secondary and 3-dimensional structures

Feng-shan, Gao; Bai, Jing; Zhang, Qiang; Chongbo, Xu; Li, Yanmin

doi:10.1016/j.gene.2012.04.038

Cited by 6 publications

(5 citation statements)

References 16 publications

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“…Nevertheless, only a very limited set of the SLA class I molecules have been well documented and reported ( Zhang N. et al, 2011 ; Fan et al, 2016 ), and the characteristics of peptide presentation for SLA class I molecules and cellular immune mechanisms have remained elusive until now. The structure and function of SLA class I complexes constructed in vitro are currently used to simulate the functions of SLA class I molecules in vivo and numerous in vitro SLA class I complexes have been constructed and different peptide–SLA-I binding assays have been suggested ( Oleksiewicz et al, 2002 ; Sylvester-Hvid et al, 2002 ; Gao et al, 2006 ; Pedersen et al, 2011 ; Zhang N. et al, 2011 ; Gao et al, 2012 ; Fan et al, 2016 ). A relative simple and rapid, in vitro refolding enzyme-linked immunosorbent assay (ELISA)-based method was able to discriminate between peptide-occupied and peptide-free SLA-I complexes based on monoclonal antibody PT85A binding ( Oleksiewicz et al, 2002 ), because the PT85A monoclonal antibody can recognize all SLA class I molecules, from outbred as well as inbred pigs, and the conformational epitope was recognized by PT85A, required the presence of the ‘correct’ peptide as well as the ‘correct’ SLA class I molecule sequence ( Lunney, 1994 ; Davis et al, 2000 ; Mosaad et al, 2006 ).…”

Section: Introductionmentioning

confidence: 99%

Specificity Characterization of SLA Class I Molecules Binding to Swine-Origin Viral Cytotoxic T Lymphocyte Epitope Peptides in Vitro

Gao

Quan

et al. 2017

Front. Microbiol.

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Swine leukocyte antigen (SLA) class I molecules play a crucial role in generating specific cellular immune responses against viruses and other intracellular pathogens. They mainly bind and present antigens of intracellular origin to circulating MHC I-restricted cytotoxic T lymphocytes (CTLs). Binding of an appropriate epitope to an SLA class I molecule is the single most selective event in antigen presentation and the first step in the killing of infected cells by CD8+ CTLs. Moreover, the antigen epitopes are strictly restricted to specific SLA molecules. In this study, we constructed SLA class I complexes in vitro comprising viral epitope peptides, the extracellular region of the SLA-1 molecules, and β2-microglobulin (β2m) using splicing overlap extension polymerase chain reaction (SOE-PCR). The protein complexes were induced and expressed in an Escherichia coli prokaryotic expression system and subsequently purified and refolded. Specific binding of seven SLA-1 proteins to one classical swine fever virus (CSFV) and four porcine reproductive and respiratory syndrome virus (PRRSV) epitope peptides was detected by enzyme-linked immunosorbent assay (ELISA)-based method. The SLA-1∗13:01, SLA-1∗11:10, and SLA-1∗11:01:02 proteins were able to bind specifically to different CTL epitopes of CSFV and PRRSV and the MHC restrictions of the five epitopes were identified. The fixed combination of Asn151Val152 residues was identified as the potentially key amino acid residues influencing the binding of viral several CTL epitope peptides to SLA-1∗13:01 and SLA-1∗04:01:01 proteins. The more flexible pocket E in the SLA-1∗13:01 protein might have fewer steric limitations and therefore be able to accommodate more residues of viral CTL epitope peptides, and may thus play a critical biochemical role in determining the peptide-binding motif of SLA-1∗13:01. Characterization of the binding specificity of peptides to SLA class I molecules provides an important basis for epitope studies of infectious diseases in swine, and for the rational development of novel porcine vaccines, as well as for detailed studies of CTL responses in pigs used as animal models.

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Section: Introductionmentioning

confidence: 99%

Specificity Characterization of SLA Class I Molecules Binding to Swine-Origin Viral Cytotoxic T Lymphocyte Epitope Peptides in Vitro

Gao

Quan

et al. 2017

Front. Microbiol.

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“…Based on human HLA-A2 crystal data, the structure of SLA-2*HB01 was predicted by using homology modeling. We noticed SLA-2*HB01 preserved some key functional sites of HLA-A2 and H-2, which indicated that SLA-2*HB01 should be crucial in binding and presenting antigenic peptides [23, 24]. In addition, SLA-2*HB01 has 85.0–93.9, 86.2–97.0 and 83.3–88.6% sequence identity to other SLA-1, SLA-2 and SLA-3 alleles, respectively.…”

Section: Discussionmentioning

confidence: 99%

Expression, purification, crystallization and preliminary X-ray diffraction analysis of swine leukocyte antigen 2 complexed with a CTL epitope AS64 derived from Asia1 serotype of foot-and-mouth disease virus

et al. 2018

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BackgroundCurrently, the structural characteristics of the swine major histocompatibility complex (MHC) class I molecule, also named swine leukocyte antigen class I (SLA-I) molecule need to be further clarified.ResultsA complex of SLA-I constituted by an SLA-2*HB01 molecule with swine β2-microglobulin and a cytotoxic T lymphocyte (CTL) epitope FMDV-AS64 (ALLRSATYY) derived from VP1 protein (residues 64–72) of Asia 1 serotype of foot-and-mouth disease virus (FMDV) was expressed, refolded, purified and crystallized. By preliminary X-ray diffraction analysis, it was shown that the diffraction resolution of the crystal was 2.4 Å and the space group belonged to P212121 with unit cell parameters a = 48.37, b = 97.75, c = 166.163 Å.ConclusionThis research will be in favor of illuminating the structural characteristics of an SLA-2 molecule associated with a CTL epitope derived from Asia1 serotype of FMDV.

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“…To achieve higher resolution, researchers have used Bessel beams to generate the illumination sheet in LSFM (68)(69)(70)(71)(72)(73)(74)(75). Although conventional LSFM has increased the speed of volumetric imaging, the use of Gaussian light sheets limits the axial resolution to 1-10 μm, trading the thickness of sectioning for the focal uniformity of the sheet.…”

Section: Resolutionmentioning

confidence: 99%

“…Although conventional LSFM has increased the speed of volumetric imaging, the use of Gaussian light sheets limits the axial resolution to 1-10 μm, trading the thickness of sectioning for the focal uniformity of the sheet. To address this shortcoming, several approaches have applied Bessel beams to generate an ultrathin light sheet of illumination (69,70,72). Scanned Bessel beams coupled to structured illumination can create light sheets thinner than 0.5 μm for subcellular imaging of microtubules, mitochondria, and mitotic spindles (69,70).…”

Section: Resolutionmentioning

confidence: 99%

“…To address this shortcoming, several approaches have applied Bessel beams to generate an ultrathin light sheet of illumination (69,70,72). Scanned Bessel beams coupled to structured illumination can create light sheets thinner than 0.5 μm for subcellular imaging of microtubules, mitochondria, and mitotic spindles (69,70). The advantage of Bessel beams relies on their extended focus; however, they are affected by high power density and out-of-focus illumination due to the presence of outer rings of the Bessel beams' structures.…”

Section: Resolutionmentioning

confidence: 99%

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Imaging, Visualization, and Computation in Developmental Biology

Cutrale

Fraser

2019

Annu. Rev. Biomed. Data Sci.

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Embryonic development is highly complex and dynamic, requiring the coordination of numerous molecular and cellular events at precise times and places. Advances in imaging technology have made it possible to follow developmental processes at cellular, tissue, and organ levels over time as they take place in the intact embryo. Parallel innovations of in vivo probes permit imaging to report on molecular, physiological, and anatomical events of embryogenesis, but the resulting multidimensional data sets pose significant challenges for extracting knowledge. In this review, we discuss recent and emerging advances in imaging technologies, in vivo labeling, and data processing that offer the greatest potential for jointly deciphering the intricate cellular dynamics and the underlying molecular mechanisms. Our discussion of the emerging area of “image-omics” highlights both the challenges of data analysis and the promise of more fully embracing computation and data science for rapidly advancing our understanding of biology.

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Construction of multiple recombinant SLA-I proteins by linking heavy chains and light chains in vitro and analyzing their secondary and 3-dimensional structures

Cited by 6 publications

References 16 publications

Specificity Characterization of SLA Class I Molecules Binding to Swine-Origin Viral Cytotoxic T Lymphocyte Epitope Peptides in Vitro

Specificity Characterization of SLA Class I Molecules Binding to Swine-Origin Viral Cytotoxic T Lymphocyte Epitope Peptides in Vitro

Expression, purification, crystallization and preliminary X-ray diffraction analysis of swine leukocyte antigen 2 complexed with a CTL epitope AS64 derived from Asia1 serotype of foot-and-mouth disease virus

Imaging, Visualization, and Computation in Developmental Biology

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