MAJOR INTEGRAL MEMBRANE-PROTEIN IMMUNOGENS OF TREPONEMA-PALLIDUM ARE PROTEOLIPIDS
MAJOR INTEGRAL MEMBRANE-PROTEIN IMMUNOGENS OF TREPONEMA-PALLIDUM ARE PROTEOLIPIDS
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DOI：
10.1128/iai.57.9.2872-2877.1989
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发表时间：
1989-09-01
影响因子：
3.1
通讯作者：
NORGARD, MV
中科院分区：
文献类型：
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作者：
CHAMBERLAIN, NR;BRANDT, ME;NORGARD, MV
梅毒螺旋体的许多主要病原体特异性免疫原最近被表征为通过用Triton X-114相分配的两亲性、完整的膜蛋白（J.D. Radolf，N. R.张伯伦，A. Clausell和M.维·诺加德Infect. Immun. 56：490-498，1988）。在本研究中，我们证明了相同的膜免疫原（指定为去污剂相蛋白[DPP]）成为放射性标记后，在体外孵育T。苍白球与各种3 H标记的脂肪酸。用单克隆抗体进行放射免疫沉淀证实，3 H标记的47千道尔顿蛋白对应于具有相同表观分子量的已充分表征的密螺旋体抗原。未能检测到3 H-标记的DPPs与红霉素孵育后证实，蛋白质酰化需要从头蛋白质合成的细菌。当密螺旋体与[3 H]肉豆蔻酸酯、[3 H]棕榈酸酯或[3 H]油酸酯孵育时，放射自显影检测到对应于DPP的放射性标记蛋白。证明T.苍白球是蛋白脂质，提供了有助于阐明其膜结合的信息，并可用于解释其非凡的免疫原性。
A number of the major pathogen-specific immunogens of Treponema pallidum were characterized recently as amphiphilic, integral membrane proteins by phase partitioning with Triton X-114 (J. D. Radolf, N. R. Chamberlain, A. Clausell, and M. V. Norgard. Infect. Immun. 56: 490-498, 1988). In the present study, we demonstrated that the same membrane immunogens (designated as detergent phase proteins [DPPs]) become radiolabeled upon in vitro incubation of T. pallidum with various 3H-labeled fatty acids. Radioimmunoprecipitation with a monoclonal antibody confirmed that the 3H-labeled 47-kilodalton protein corresponded to the well-characterized treponemal antigen with the identical apparent molecular mass. Failure to detect 3H-labeled DPPs following incubation with erythromycin confirmed that protein acylation required de novo protein synthesis by the bacteria. When treponemes were incubated with [3H]myristate, [3H]palmitate, or [3H]oleate, radiolabeled proteins corresponding to the DPPs were detected upon autoradiography. Demonstration that a number of the abundant membrane immunogens of T. pallidum are proteolipids provides information to help clarify their membrane association(s) and may serve to explain their extraordinary immunogenicity.