MHCⅠ类分子BF2*0201递呈ALV-J表位结构和递呈机制解析

doi:10.11843/j.issn.0366-6964.2024.07.031

摘要/Abstract

摘要：

MHC B2单倍型鸡对禽白血病病毒感染有较强抗性，这种抗病性可能与被MHC分子递呈的表位刺激活化的T细胞产生的免疫应答有关。本研究拟阐明MHC Ⅰ类分子BF2*0201递呈ALV-J表位结构和递呈机制。利用凝胶过滤层析筛选出能与MHC Ⅰ稳定结合的肽。利用晶体初筛试剂盒在体外培育肽和MHC Ⅰ的二元复合物晶体，并利用COOT、CCP4和PYMOL等软件对该晶体结构进行解析和分析。在这项研究中，作者发现二个与BF2*0201稳定结合的ALV-J CTL表位FVDFANRLI和SALQAFREV。得到一个优质晶体BF2*0201-SALQAFREV(SV9)，其生长条件为1.0 mol ·L^-1 丁二酸(pH 7.0)，0.1 mol ·L^-1 HEPES (pH 7.0)，1%聚乙二醇单甲醚2000。解析高分辨率(1.72 Å)的BF2*0201-SV9复合物结构后发现BF2*0201抗原结合槽整体呈现弱负电性。保守的残基多位于抗原结合槽的两端，特异性氨基酸赋予BF2*0201中等大小的抗原结合槽。B、C和F口袋起着较重要的锚定作用，口袋残基和水分子与抗原多肽形成氢键作用，将其固定在抗原结合槽中。SV9表位构象中突出的P4-Gln和P7-Arg残基的构象特征表明该位点有被特异性TCR潜在识别的特性。BF2*0201-SV9复合物晶体结构特征有助于阐明B2单倍型MHCⅠ类分子BF2*0201递呈ALV-J表位的机制，有助于解释B2单倍型鸡对ALV存在抗性的原因，为抗病育种工作奠定理论基础。

关键词: MHC, BF2*0201, ALV-J, 递呈机制, 晶体结构

Abstract:

MHC B2 haplotype chickens are highly resistant to avian leukosis virus infection, and this resistance may be related to the activated T cell immune response stimulated by MHC molecule presented epitopes. This study aims to elucidate the structure and mechanism of ALV-J epitope presentation by the MHC class Ⅰ molecule BF2*0201. Peptides that can bind stably to MHC Ⅰ were screened by gel filtration chromatography. The complex crystals of the peptide and MHC Ⅰ were grown in vitro using a crystal primary screening kit, and the crystal structure was resolved and analyzed using COOT, CCP4 and PYMOL. In this study, we identified two ALV-J CTL epitopes FVDFANRLI and SALQAFREV that bind stably to BF2*0201. We obtained a high quality crystal BF2*0201-SALQAFREV (SV9) grown under the conditions of 1.0 mol ·L^-1 succinic acid (pH 7.0), 0.1 mol ·L^-1 HEPES (pH 7.0), and 1% polyethylene glycol monomethyl ether 2000. We resolved the structure of the high-resolution (1.72 Å) BF2*0201-SV9 complex and found that the BF2*0201 antigen-binding groove exhibits weak electronegativity overall. Conserved residues were located at both ends of the antigen-binding groove, and specific amino acids gave BF2*0201 a medium-sized peptide-binding groove. The B, C, and F pockets played an important anchoring role. The residues in pockets and water molecules formed hydrogen bonds with the antigenic polypeptide to anchor it to the peptide-binding groove, and the antigenic polypeptide was in an "M"-shaped conformation. The conformational features of the prominent Gln-4 and Arg-7 residues in the SV9 epitope conformation suggested potential recognition of this site by specific TCRs. The crystal structure of BF2*0201-SV9 complex can help to elucidate the mechanism of ALV-J epitope presented by BF2*0201, which can help to explain the reason for the resistance of B2 haplotype chickens to ALV, and lay a theoretical foundation for the breeding of resistance to the disease.

Key words: MHC, BF2*0201, ALV-J, epitope presentation, crystal structure

中图分类号:

S852.43

贾玉生, 李易霖, 马露露, 廖明, 代曼曼. MHCⅠ类分子BF2*0201递呈ALV-J表位结构和递呈机制解析[J]. 畜牧兽医学报, 2024, 55(7): 3132-3142.

Yusheng JIA, Yilin LI, Lulu MA, Ming LIAO, Manman DAI. Structure and Mechanism of ALV-J Epitope Presented by MHC Class Ⅰ Molecule BF2*0201[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(7): 3132-3142.

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参数 Parameter	BF20201-SV9的数值 Value in BF20201-SV9
数据处理 Data processing
Space group	P1211
Cell parameters (Å)	86.614 115.584 94.107 90 111.158 90
Resolution range (Å)	43.06~1.72 (1.781~1.72)
Total reflections	354 688 (34 360)
Unique reflections	180 646 (17 706)
Completeness (%)	98.68 (97.06)
R_merge	0.042 48 (0.314 4)
重修 Refinement
R_work	0.183 1 (0.254 4)
R_free	0.215 0 (0.281 8)
RMSD
Bonds (Å)	0.010
Angles (°)	1.61
Average B factor	21.90
拉氏图质量 Ramachandran plot quality
Most favored (%)	98.93

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