Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Aichi 470-1192, Japan.
Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Aichi 470-1192, Japan.
Biochem Biophys Res Commun. 2014 Jan 10;443(2):422-7. doi: 10.1016/j.bbrc.2013.11.096. Epub 2013 Dec 2.
The major histocompatibility complex (MHC) class I-related molecule, MR1, is highly conserved in mammals and can present bacteria-derived vitamin B metabolites to mucosal-associated invariant T (MAIT) cells, possibly having important defense function in the microbial infection. MR1B is a splice variant of MR1 and possesses an intriguing domain structure with only two extracellular domains resembling some NKG2D ligand molecules. Thus far, cell surface expression of MR1B could not be analyzed with flow cytometry due to a lack of appropriate antibodies reactive with MR1B. Here we clarified the expression of MR1B recombinant protein on the cell surface of the transfected cells by flow cytometry analyses using the antiserum against MR1. Consistently, MR1B tagged with FLAG peptide at the N-terminus also could be detected with anti-FLAG monoclonal antibodies. Our result showed that MR1B can be recognized on the cell surface by macromolecules such as antibodies, indicating its potential of interaction with certain receptor(s). We discuss possibility of interaction of MR1B and/or the full-length MR1 with some receptor(s) other than αβ T cell receptor (TCR) of MAIT cells based on the highly conserved characteristic residues of the ligand-binding domains of MR1 and its MAIT cells αβTCR footprints.
主要组织相容性复合体(MHC)I 类相关分子 MR1 在哺乳动物中高度保守,可将细菌衍生的维生素 B 代谢物呈递给黏膜相关不变 T(MAIT)细胞,可能在微生物感染中具有重要的防御功能。MR1B 是 MR1 的剪接变体,具有引人注目的结构域,仅具有两个类似于某些 NKG2D 配体分子的细胞外结构域。到目前为止,由于缺乏与 MR1B 反应的合适抗体,无法通过流式细胞术分析 MR1B 的细胞表面表达。在这里,我们使用针对 MR1 的抗血清通过流式细胞术分析澄清了转染细胞表面上 MR1B 重组蛋白的表达。一致地,在 N 末端带有 FLAG 肽标签的 MR1B 也可以用抗 FLAG 单克隆抗体检测到。我们的结果表明,MR1B 可以被抗体等大分子识别到细胞表面,表明其与某些受体(s)相互作用的潜力。我们讨论了基于 MR1 的配体结合域和 MAIT 细胞的 αβTCR 足迹的高度保守特征残基,MR1B 和/或全长 MR1 与 MAIT 细胞的 αβTCR 以外的某些受体(s)相互作用的可能性。