THE CELL ATTACHMENT SITE ON FOOT-AND-MOUTH-DISEASE VIRUS INCLUDES THE AMINO-ACID SEQUENCE RGD (ARGININE-GLYCINE-ASPARTIC ACID)
THE CELL ATTACHMENT SITE ON FOOT-AND-MOUTH-DISEASE VIRUS INCLUDES THE AMINO-ACID SEQUENCE RGD (ARGININE-GLYCINE-ASPARTIC ACID)
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DOI：
10.1099/0022-1317-70-3-625
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发表时间：
1989-03-01
影响因子：
3.8
通讯作者：
BROWN, F
中科院分区：
文献类型：
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作者：
FOX, G;PARRY, NR;BROWN, F
口蹄疫病毒(FMDV)VP1蛋白中的精氨酸-甘氨酸-天冬氨酸(RGD)氨基酸序列虽然位于135-160位氨基酸之间的免疫显性高变区，但在蛋白质中高度保守。已知含有RGD的蛋白在促进几种不同系统中的细胞附着方面起重要作用，我们在此报告含有该序列的合成肽能够抑制病毒与婴儿仓鼠肾(BHK)细胞的附着。抑制作用呈剂量依赖关系，去除多肽后可逆转。与相同高变区的一部分相对应但不包含RGD序列的合成肽在相同条件下不抑制病毒附着。在EL ISA试验中，针对VP1 RGD区的抗体阻断了病毒与BHK细胞的附着，而通过阻止细胞附着来中和病毒的中和性单抗则被含有RGD的多肽阻断。用蛋白水解酶原位切割病毒VP1的C末端区域可减少细胞黏附，与该区域相对应的多肽的抗血清也能抑制病毒与BHK细胞的黏附。这些结果表明，BHK细胞的口蹄疫病毒的细胞附着部位是由145~147位氨基酸序列和VP1 C末端区域(203~213位)氨基酸组成的。
The amino acid sequence RGD (arginine-glycine-aspartic acid) is highly conserved in the VP1 protein of foot-and-mouth disease virus (FMDV), despite being situated in the immunodominant hypervariable region between amino acids 135 and 160. RGD-containing proteins are known to be important in promoting cell attachment in several different systems, and we report here that synthetic peptides containing this sequence are able to inhibit attachment of the virus to baby hamster kidney (BHK) cells. Inhibition was dose-dependent and could be reversed on removal of the peptide. A synthetic peptide corresponding to a portion of the same hypervariable region but not containing the RGD sequence did not inhibit virus attachment under the same conditions. Antibody against the RGD region of VP1 blocked attachment of the virus to BHK cells, and neutralizing monoclonal antibodies, which neutralize virus by preventing cell attachment, were blocked by RGD-containing peptides from binding virus in an ELISA test. Cleavage of the C-terminal region of virus VP1 in situ with proteolytic enzymes reduced cell attachment, and antiserum against a peptide corresponding to this region was also able to inhibit attachment of virus to BHK cells. These results indicate that the amino acid sequence RGD at positions 145 to 147 and amino acids from the C-terminal region of VP1 (positions 203 to 213) contribute to the cell attachment site on FMDV for BHK cells.