Shoemaker C, Gross A, Gebremichael A, Harn D
Department of Tropical Public Health, Harvard School of Public Health, Boston, MA 02115.
Proc Natl Acad Sci U S A. 1992 Mar 1;89(5):1842-6. doi: 10.1073/pnas.89.5.1842.
M.1 monoclonal antibody has previously been shown to passively transfer partial resistance to schistosome infection within mice and to recognize a 28-kDa antigen that has peptide sequence homology with triose-phosphate isomerase (TPI; D-glyceraldehyde-3-phosphate ketol-isomerase, EC 5.3.1.1). We have now isolated the complete coding DNA for Schistosoma mansoni TPI and confirmed that this cDNA encodes the 28-kDa antigen recognized by M.1. The predicted translation product has strong homology with other TPIs, particularly from higher eukaryotes, and the sequence homology is greatest in regions known to form the active site. The complete coding DNA has been expressed within an Escherichia coli host to produce high levels of soluble, recombinant S. mansoni TPI protein. The product is recognized and purified by the M.1 antibody and is a functional TPI with an intrinsic specific activity comparable to that of rabbit and yeast TPI.
先前已证明M.1单克隆抗体可在小鼠体内被动转移对血吸虫感染的部分抗性，并识别一种28 kDa的抗原，该抗原与磷酸丙糖异构酶（TPI；D-甘油醛-3-磷酸酮醇异构酶，EC 5.3.1.1）具有肽序列同源性。我们现已分离出曼氏血吸虫TPI的完整编码DNA，并证实该cDNA编码M.1识别的28 kDa抗原。预测的翻译产物与其他TPI具有高度同源性，特别是来自高等真核生物的TPI，并且在已知形成活性位点的区域序列同源性最高。完整的编码DNA已在大肠杆菌宿主中表达，以产生高水平的可溶性重组曼氏血吸虫TPI蛋白。该产物可被M.1抗体识别和纯化，并且是一种具有内在比活性的功能性TPI，其内在比活性与兔和酵母TPI相当。
