Vanbuskirk A, Crump B L, Margoliash E, Pierce S K
Department of Biochemistry, Molecular Biology and Cell Biology, Northwestern University, Evanston, Illinois 60208.
J Exp Med. 1989 Dec 1;170(6):1799-809. doi: 10.1084/jem.170.6.1799.
The T cell recognition of globular protein antigens requires the processing and presentation of the antigen by Ia-expressing APCs. Processing is believed to involve the uptake of antigen into an acidic compartment where proteolysis occurs. The resulting peptides containing the T cell antigenic determinant are associated with Ia and presented at the cell surface to the specific T cells. The mechanisms by which antigenic peptides become associated with Ia is not known. We previously described a peptide binding protein of 72/74 x 10(3) Mr (PBP72/74) that plays a role in antigen presentation as shown by the ability of an antiserum raised in rabbits to affinity-purified PBP72/74 to block presentation of cytochrome c to a cytochrome c-specific T cell hybrid. Here we show that PBP72/74 is recognized by mAbs specific for members of the HSP70 family of proteins. In Western blots PBP72/74 is bound by mAb 7.10, specific for an evolutionarily conserved epitope of HSP proteins and by mAb N27, specific for both the constitutively expressed and inducible 72/73 x 10(3) Mr HSP70 proteins. In addition, PBP72/74 shares a second common feature of the HSP proteins, that of binding to ATP. Indeed, ATP causes the release of PBP72/74 from binding to a peptide fragment of cytochrome c (Pc 81-104) and PBP72/74 can be eluted from ATP columns by Pc 81-104. Finally, a portion of PBP72/74 is shown to be present on B cell surfaces by immunofluorescence staining. Thus, it appears that characteristics of the heat shock proteins are shared by a protein playing a role in antigen presentation, suggesting some commonality in function.
T细胞对球状蛋白抗原的识别需要由表达Ia的抗原呈递细胞(APC)对抗原进行加工和呈递。据信加工过程涉及抗原被摄取到一个发生蛋白水解的酸性区室中。产生的含有T细胞抗原决定簇的肽与Ia相关联,并呈递到细胞表面给特异性T细胞。抗原肽与Ia相关联的机制尚不清楚。我们先前描述了一种分子量为72/74×10³的肽结合蛋白(PBP72/74),它在抗原呈递中发挥作用,这可通过用兔抗亲和纯化的PBP72/74血清阻断细胞色素c呈递给细胞色素c特异性T细胞杂交体的能力得以证明。在此我们表明PBP72/74可被针对热休克蛋白70(HSP70)家族成员的单克隆抗体(mAb)识别。在蛋白质印迹中,PBP72/74被对HSP蛋白进化保守表位特异的mAb 7.10以及对组成型表达和诱导型72/73×10³的HSP70蛋白均特异的mAb N27所结合。此外,PBP72/74具有HSP蛋白的第二个共同特征,即与ATP结合。实际上,ATP导致PBP72/74从与细胞色素c的肽片段(Pc 81 - 104)的结合中释放出来,并且PBP72/74可被Pc 81 - 104从ATP柱上洗脱下来。最后,通过免疫荧光染色显示一部分PBP72/74存在于B细胞表面。因此,在抗原呈递中发挥作用的一种蛋白似乎具有热休克蛋白的特征,提示在功能上存在一些共性。