Plasmodium vivax merozoite surface proteins-3β and-3γ share structural similarities with P. vivax merozoite surface protein-3α and define a new gene family
Plasmodium vivax merozoite surface proteins-3β and-3γ share structural similarities with P. vivax merozoite surface protein-3α and define a new gene family
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DOI：
10.1016/s0166-6851(01)00267-5
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发表时间：
2001-06-01
影响因子：
1.5
通讯作者：
Barnwell, JW
中科院分区：
文献类型：
--
作者：
Galinski, MR;Ingravallo, P;Barnwell, JW
报道了与PvMSP 3相关的两个间日疟原虫裂殖子表面蛋白的编码基因[1]。在间日疟原虫λ gt 11克隆5.4中鉴定了这些基因之一，该基因是通过用赛米里猴抗血清进行免疫筛选而选择的。将该克隆的插入DNA用作探针以从间日疟原虫λ DASH基因组（g）DNA文库中分离完整基因。重组5.4和随后的融合蛋白的抗体通过间接免疫荧光测定（IFA）在分节的裂殖子和游离的完整裂殖子上产生圆周表面荧光的模式，并通过蛋白质免疫印迹识别125 kDa蛋白。然而，该基因编码的蛋白质的计算大小为75 677 Da，和3 '和5 ' RACE分析，以确认该基因及其编码区的大小。第二个相关的间日疟原虫基因是通过与一个直链诺氏疟原虫基因片段杂交而分离的。所有三种相关的间日疟原虫基因的编码蛋白具有推定的信号肽，即含有> 20%由带电区域结合的丙氨酸残基的大的中心结构域，预测其形成具有七肽重复卷曲螺旋结构的α-螺旋，并且不具有可以将它们锚到裂殖子表面的疏水区域。尽管三种编码蛋白质之间氨基酸比对的总体同一性较低（< 40%），但共有的预测结构特征和基序表明它们是种内家族的成员，我们将其命名为PvMSP-3家族，其中报道的成员为Pvmsp-3 α、Pvmsp-3 β和Pvmsp-3 γ。我们进一步证明，这个家庭也包括相关的蛋白质从P，诺氏疟原虫和恶性疟原虫。(C)2001 Elsevier Science B. V.保留所有权利。
The genes encoding two merozoite surface proteins of Plasmodium vivax that are related to PvMSP3 [1] are reported. One of these genes was identified within P. vivax, lambda gt11 clone 5.4, which was selected by immunoscreening with a Saimiri monkey antiserum. The insert DNA of this clone was used as a probe to isolate the complete gene from a P. vivax lambda DASH genomic (g) DNA library. Antibodies to recombinant 5.4 and subsequent fusion proteins produce a pattern of circumferential surface fluorescence by indirect immunofluorescence assays (IFA) on segmented schizonts and free intact merozoites, and recognize a 125 kDa protein via western immunoblots. The gene, however, encodes a protein with a calculated size of 75 677 Da, and 3 ' and 5 ' RACE analyses were employed to confirm the size of the gene and its coding region. The second related P. vivax gene was isolated by hybridization of a fragment of an orthologous P. knowlesi gene. The encoded proteins of all three related P. vivax genes have putative signal peptides, large central domains that contain > 20% alanine residues bound by charged regions, are predicted to form alpha -helices with heptad repeat coiled-coil structures, and do not have a hydrophobic region that could anchor them to the surface of the merozoite. Although the overall identity in amino acid alignment among the three encoded proteins is low ( < 40%) the shared predicted structural features and motifs indicate that they are members of an intra-species family, which we are designating as the PvMSP-3 family with the reported members being Pvmsp-3 alpha, Pvmsp-3 beta, and Pvmsp-3 gamma. We further demonstrate that this family also includes related proteins from P, knowlesi and P. falciparum. (C) 2001 Elsevier Science B.V. All rights reserved.