Expression, refolding and preliminary X-ray crystallographic analysis of equine MHC class I molecule complexed with an EIAV-Env CTL epitope.
Abstract: In order to clarify the structure and the peptide-presentation characteristics of the equine major histocompatibility complex (MHC) class I molecule, a complex of equine MHC class I molecule (ELA-A1 haplotype, 7-6 allele) with mouse β(2)-microglobulin and the cytotoxic T lymphocyte (CTL) epitope Env-RW12 (RVEDVTNTAEYW) derived from equine infectious anaemia virus (EIAV) envelope protein (residues 195-206) was refolded and crystallized. The crystal, which belonged to space group P2(1), diffracted to 2.3 Å resolution and had unit-cell parameters a = 82.5, b = 71.4, c = 99.8 Å, β = 102.9°. The crystal structure contained two molecules in the asymmetric unit. These results should help to determine the first equine MHC class I molecule structure presenting an EIAV CTL epitope.
© 2012 International Union of Crystallography. All rights reserved.
Publication Date: 2011-12-24 PubMed ID: 22232164PubMed Central: PMC3253827DOI: 10.1107/S1744309111038139Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
This study attempted to reveal the structure and peptide-presenting properties of the equine major histocompatibility complex (MHC) class I molecule, by creating a complex with a mouse protein and a cytotoxic cell epitope from the equine infectious anaemia virus (EIAV).
Understanding the objectives
- The primary aim of this research was to understand the characteristics of the equine MHC class I molecule, particularly in terms of its structure and its capabilities in presenting peptides.
- Precisely, they wanted to investigate how this molecule interacts with the cytotoxic T lymphocyte (CTL) epitope derived from EIAV, a virus that causes infectious anaemia in horses.
Methodology applied in the study
- To achieve the research objectives, a complex was created involving the equine MHC class I molecule, mouse β(2)-microglobulin, and the CTL epitope Env-RW12 derived from EIAV envelope protein.
- This complex was then refolded and crystallized for further analysis.
- The resultant crystal was categorized into the space group P2(1), with specific unit-cell parameters.
- X-ray crystallographic analysis was conducted to achieve a resolution of 2.3 Å. This means that the researchers were able to observe in detail, up to 2.3 angstrom (a unit of length used specifically in the field of crystallography).
Outcome of the research
- It was found that the crystal structure contained two molecules in the asymmetric unit, a noteworthy observation in the study.
- This finding and the overall results from the study provide valuable insights needed to determine the first known structure of an equine MHC class I molecule which presents an EIAV CTL epitope.
- From a broader perspective, the research contributes to the existing body of knowledge regarding the role of MHC class I molecules in health and disease, and particularly its significance concerning viral infections in equines.
Cite This Article
APA
Yao S, Qi J, Liu J, Chen R, Pan X, Li X, Gao F, Xia C.
(2011).
Expression, refolding and preliminary X-ray crystallographic analysis of equine MHC class I molecule complexed with an EIAV-Env CTL epitope.
Acta Crystallogr Sect F Struct Biol Cryst Commun, 68(Pt 1), 20-23.
https://doi.org/10.1107/S1744309111038139 Publication
Researcher Affiliations
- Department of Microbiology and Immunology, College of Veterinary Medicine, China Agricultural University, Yuanmingyuan Xi Lu No. 2, Haidian District, Beijing 100094, People's Republic of China.
MeSH Terms
- Amino Acid Sequence
- Animals
- Crystallography, X-Ray
- Epitopes, T-Lymphocyte / chemistry
- Epitopes, T-Lymphocyte / immunology
- Epitopes, T-Lymphocyte / metabolism
- Gene Expression
- Gene Products, env / chemistry
- Gene Products, env / immunology
- Histocompatibility Antigens Class I / chemistry
- Histocompatibility Antigens Class I / immunology
- Histocompatibility Antigens Class I / metabolism
- Horses / immunology
- Horses / metabolism
- Infectious Anemia Virus, Equine / chemistry
- Infectious Anemia Virus, Equine / immunology
- Molecular Sequence Data
- Protein Folding
- T-Lymphocytes, Cytotoxic / chemistry
- T-Lymphocytes, Cytotoxic / immunology
- T-Lymphocytes, Cytotoxic / metabolism
References
This article includes 20 references
- Achour A, Persson K, Harris RA, Sundbäck J, Sentman CL, Lindqvist Y, Schneider G, Kärre K. The crystal structure of H-2Dd MHC class I complexed with the HIV-1-derived peptide P18-I10 at 2.4 A resolution: implications for T cell and NK cell recognition.. Immunity 1998 Aug;9(2):199-208.
- Ansari HA, Hediger R, Fries R, Stranzinger G. Chromosomal localization of the major histocompatibility complex of the horse (ELA) by in situ hybridization.. Immunogenetics 1988;28(5):362-4.
- Bjorkman PJ, Saper MA, Samraoui B, Bennett WS, Strominger JL, Wiley DC. Structure of the human class I histocompatibility antigen, HLA-A2.. Nature 1987 Oct 8-14;329(6139):506-12.
- Chung C, Leib SR, Fraser DG, Ellis SA, McGuire TC. Novel classical MHC class I alleles identified in horses by sequencing clones of reverse transcription-PCR products.. Eur J Immunogenet 2003 Dec;30(6):387-96.
- Cohen J. HIV/AIDS in China. Vaccine development with a distinctly Chinese flavor.. Science 2004 Jun 4;304(5676):1437.
- Diederichs K, Karplus PA. Improved R-factors for diffraction data analysis in macromolecular crystallography.. Nat Struct Biol 1997 Apr;4(4):269-75.
- Garboczi DN, Ghosh P, Utz U, Fan QR, Biddison WE, Wiley DC. Structure of the complex between human T-cell receptor, viral peptide and HLA-A2.. Nature 1996 Nov 14;384(6605):134-41.
- Kantardjieff KA, Rupp B. Matthews coefficient probabilities: Improved estimates for unit cell contents of proteins, DNA, and protein-nucleic acid complex crystals.. Protein Sci 2003 Sep;12(9):1865-71.
- Lazary S, Antczak DF, Bailey E, Bell TK, Bernoco D, Byrns G, McClure JJ. Joint Report of the Fifth International Workshop on Lymphocyte Alloantigens of the Horse, Baton Rouge, Louisiana, 31 October-1 November 1987.. Anim Genet 1988;19(4):447-56.
- Li X, Liu J, Qi J, Gao F, Li Q, Li X, Zhang N, Xia C, Gao GF. Two distinct conformations of a rinderpest virus epitope presented by bovine major histocompatibility complex class I N*01801: a host strategy to present featured peptides.. J Virol 2011 Jun;85(12):6038-48.
- Liu J, Wu P, Gao F, Qi J, Kawana-Tachikawa A, Xie J, Vavricka CJ, Iwamoto A, Li T, Gao GF. Novel immunodominant peptide presentation strategy: a featured HLA-A*2402-restricted cytotoxic T-lymphocyte epitope stabilized by intrachain hydrogen bonds from severe acute respiratory syndrome coronavirus nucleocapsid protein.. J Virol 2010 Nov;84(22):11849-57.
- Matthews BW. Solvent content of protein crystals.. J Mol Biol 1968 Apr 28;33(2):491-7.
- Mealey RH, Lee JH, Leib SR, Littke MH, McGuire TC. A single amino acid difference within the alpha-2 domain of two naturally occurring equine MHC class I molecules alters the recognition of Gag and Rev epitopes by equine infectious anemia virus-specific CTL.. J Immunol 2006 Nov 15;177(10):7377-90.
- Mealey RH, Sharif A, Ellis SA, Littke MH, Leib SR, McGuire TC. Early detection of dominant Env-specific and subdominant Gag-specific CD8+ lymphocytes in equine infectious anemia virus-infected horses using major histocompatibility complex class I/peptide tetrameric complexes.. Virology 2005 Aug 15;339(1):110-26.
- Mealey RH, Zhang B, Leib SR, Littke MH, McGuire TC. Epitope specificity is critical for high and moderate avidity cytotoxic T lymphocytes associated with control of viral load and clinical disease in horses with equine infectious anemia virus.. Virology 2003 Sep 1;313(2):537-52.
- Otwinowski Z, Minor W. Processing of X-ray diffraction data collected in oscillation mode.. Methods Enzymol 1997;276:307-26.
- Raiola A, Camardella L, Giovane A, Mattei B, De Lorenzo G, Cervone F, Bellincampi D. Two Arabidopsis thaliana genes encode functional pectin methylesterase inhibitors.. FEBS Lett 2004 Jan 16;557(1-3):199-203.
- Turner SJ, Kedzierska K, Komodromou H, La Gruta NL, Dunstone MA, Webb AI, Webby R, Walden H, Xie W, McCluskey J, Purcell AW, Rossjohn J, Doherty PC. Lack of prominent peptide-major histocompatibility complex features limits repertoire diversity in virus-specific CD8+ T cell populations.. Nat Immunol 2005 Apr;6(4):382-9.
- Yewdell JW, Reits E, Neefjes J. Making sense of mass destruction: quantitating MHC class I antigen presentation.. Nat Rev Immunol 2003 Dec;3(12):952-61.
- Zhang J, Chen Y, Gao F, Chen W, Qi J, Xia C. Complex assembly, crystallization and preliminary X-ray crystallographic studies of duck MHC class I molecule.. Acta Crystallogr Sect F Struct Biol Cryst Commun 2010 Jan 1;66(Pt 1):99-101.
Citations
This article has been cited 2 times.- Yuan H, Chen R, Liu Y, Tariq M, Sun Y, Xia C. Crystallization and preliminary crystallographic studies of the complement 1qA globular domain from zebrafish, Dare-C1qAgD. Acta Crystallogr F Struct Biol Commun 2014 Jul;70(Pt 7):911-4.
- Wang Z, Chen R, Tariq M, Jiang B, Chen Z, Xia C. Complex assembly, crystallization and preliminary X-ray crystallographic analysis of the bovine CD8αα-BoLA-2*02201 complex. Acta Crystallogr F Struct Biol Commun 2014 Jun;70(Pt 6):742-6.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
