Olsen A C, Pedersen L O, Hansen A S, Nissen M H, Olsen M, Hansen P R, Holm A, Buus S
Institute for Medical Microbiology and Immunology, Medical Faculty, University of Copenhagen, Denmark.
Eur J Immunol. 1994 Feb;24(2):385-92. doi: 10.1002/eji.1830240218.
A direct and sensitive biochemical assay to measure the interaction in solution between peptides and affinity-purified major histocompatibility complex (MHC) class I molecules has been generated. Specific binding reflecting the known class I restriction of cytotoxic T cell responses was obtained. Adding an excess of beta 2-microglobulin (beta 2m) significantly increased the rate of peptide association, but it did not affect the rate of dissociation. Binding was complicated by a rapid and apparently irreversible loss of functional MHC class I at 37 degrees C which might limit the life span of empty MHC class I thereby preventing the inadvertent exchange of peptides at the target cell surface. All class I molecules tested bound peptides of the canonical octa- to nona-meric length. However, one class I molecule, Kk, also bound peptides, which were much longer suggesting that the preference of class I molecules for short epitopes is not absolute and may be caused by factors other than the peptide-MHC class I binding event itself.
已开发出一种直接且灵敏的生化检测方法，用于测量肽与亲和纯化的主要组织相容性复合体（MHC）I类分子在溶液中的相互作用。获得了反映细胞毒性T细胞反应已知I类限制的特异性结合。添加过量的β2-微球蛋白（β2m）显著提高了肽结合速率，但不影响解离速率。在37℃下功能性MHC I类分子快速且明显不可逆的丧失使结合变得复杂，这可能会限制空MHC I类分子的寿命，从而防止靶细胞表面肽的意外交换。所有测试的I类分子都结合了典型的八聚体至九聚体长度的肽。然而，一种I类分子Kk也结合了长得多的肽，这表明I类分子对短表位的偏好并非绝对，可能是由肽-MHC I类结合事件本身以外的因素引起的。
