Identification, design and synthesis of tubulin-derived peptides as novel hyaluronan mimetic ligands for the receptor for hyaluronan-mediated motility (RHAMM/HMMR).
Identification, design and synthesis of tubulin-derived peptides as novel hyaluronan mimetic ligands for the receptor for hyaluronan-mediated motility (RHAMM/HMMR).
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DOI：
10.1039/c5ib00222b
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发表时间：
2015-12
期刊：
影响因子：
--
通讯作者：
Luyt LG
中科院分区：
文献类型：
--
作者：
Esguerra KV;Tolg C;Akentieva N;Price M;Cho CF;Lewis JD;McCarthy JB;Turley EA;Luyt LG
细胞外基质组分透明质酸（HA）的片段促进组织炎症、纤维化和肿瘤进展。HA片段通过HA受体（包括CD 44、LYVE 1、TLR 2、4和透明质酸介导的运动性受体（RHAMM/HMMR））起作用。RHAMM是一种多功能蛋白，在细胞运动和增殖中具有细胞内和细胞外作用。细胞外RHAMM直接结合HA片段，而细胞内RHAMM直接结合ERK 1和微管蛋白。HA和微管蛋白（s-微管蛋白）的区域都是阴离子的，并结合到配偶体蛋白中富含碱性氨基酸的区域，例如RHAMM的HA和微管蛋白结合区域。我们使用这作为开发基于生物信息学和SPR（表面等离子体共振）的筛选以鉴定高亲和力阴离子RHAMM肽配体的基本原理。基于s-微管蛋白同种型的羧基末端尾部序列制备12-mer肽的文库，并使用SPR测定它们结合重组RHAMM的HA/微管蛋白结合区的能力。这种方法导致三个12-mer肽的分离与RHAMM的纳摩尔亲和力。这些肽选择性结合RHAMM，但不结合CD 44或TLR 2，4，并阻断RHAMM：HA相互作用。此外，PC 3 MLN 4前列腺癌细胞对荧光素肽的摄取被RHAMM mAb阻断，但不被CD 44 mAb阻断。这些肽还降低了前列腺癌细胞降解I型胶原蛋白的能力。这些新的HA肽模拟物对RHAMM的选择性表明它们作为HA模拟成像和HA促进的疾病的治疗剂的发展潜力。
Fragments of the extracellular matrix component hyaluronan (HA) promote tissue inflammation, fibrosis and tumor progression. HA fragments act through HA receptors including CD44, LYVE1, TLR2,4 and the receptor for hyaluronan mediated motility (RHAMM/HMMR). RHAMM is a multifunctional protein with both intracellular and extracellular roles in cell motility and proliferation. Extracellular RHAMM binds directly to HA fragments while intracellular RHAMM binds directly to ERK1 and tubulin. Both HA and regions of tubulin (s-tubulin) are anionic and bind to basic amino acid-rich regions in partner proteins, such as in HA and tubulin binding regions of RHAMM. We used this as a rationale for developing bioinformatics and SPR (surface plasmon resonance) based screening to identify high affinity anionic RHAMM peptide ligands. A library of 12-mer peptides was prepared based on the carboxyl terminal tail sequence of s-tubulin isoforms and assayed for their ability to bind to the HA/tubulin binding region of recombinant RHAMM using SPR. This approach resulted in the isolation of three 12-mer peptides with nanomolar affinity for RHAMM. These peptides bound selectively to RHAMM but not to CD44 or TLR2,4 and blocked RHAMM:HA interactions. Furthermore, fluorescein-peptide uptake by PC3MLN4 prostate cancer cells was blocked by RHAMM mAb but not by CD44 mAb. These peptides also reduced the ability of prostate cancer cells to degrade collagen type I. The selectivity of these novel HA peptide mimics for RHAMM suggest their potential for development as HA mimetic imaging and therapeutic agents for HA-promoted disease.