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Substitution of the Echistatin Amino Acid Motif RGDD with KGDW Enhances Inhibition of Platelet Aggregation and Thrombogenesis

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Abstract

Disintegrins are a family of small proteins found in snake venom. These proteins are of great biomedical importance due to their binding affinities with different kinds of integrins, which results in the inhibition of platelet aggregation, the adhesion of cancer cells, and the induction of signal transduction pathways. Disintegrins are multi-functional due to their lower integrin selectivity. To increase their binding specificity with platelets, we modified the gene sequence encoding one kind of disintegrins, echistatin (Ech). DNA recombination technology was used to change the Ech amino acid sequence RGDD to KGDW, which are located at the 24th–27th location site of Ech, thereby creating a mutant protein echistatin (E-KW). The E-KW was expressed, isolated and purified using molecular biological methods. ADP-induced platelet aggregation in human platelet-rich plasma was inhibited by E-KW at 45 nM and by Ech at an IC50 of 140 nM (P < 0.05). E-KW was also more effective than wild-type Ech in inhibiting thrombus formation (12.8 ± 3.2 vs. 27.9 ± 4.1, P < 0.05). Our studies revealed that, compared to wild-type Ech, E-KW had stronger binding specificity to the glucoprotein receptors on platelet membranes and was more effective in inhibiting platelet aggregation and thrombogenesis.

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Acknowledgments

We thank Dong-dong Sun for his involvement in the initiation of experiments. This research was supported by grants from the National Natural Science Foundation of China (81101895), Program for the Top Young Academic Leaders of Higher Learning Institutions and Program for the Scientific Activities of Selected Returned Overseas Professionals in Shanxi Province.

Conflict of interests

The authors declared that they have no conflicts of interest to this work.

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Shanxi Medical University, Xinjian South Road 56, Taiyuan, 030001, Shanxi, China

    Li-Jun Yang, Bo Niu, Dong Zhang & Tao Yang

  2. Key Laboratory of Cellular Physiology, Ministry of Education, Shanxi Medical University, Taiyuan, 030001, Shanxi, China

    Li-Jun Yang & Tao Yang

  3. Capital Institute of Pediatrics, Beijing, 100020, China

    Bo Niu

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  1. Li-Jun Yang
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  2. Bo Niu
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  4. Tao Yang
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Correspondence to Tao Yang.

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Yang, LJ., Niu, B., Zhang, D. et al. Substitution of the Echistatin Amino Acid Motif RGDD with KGDW Enhances Inhibition of Platelet Aggregation and Thrombogenesis. Int J Pept Res Ther 21, 451–458 (2015). https://doi.org/10.1007/s10989-015-9475-7

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  • DOI: https://doi.org/10.1007/s10989-015-9475-7

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