Collagen-binding growth factors: Production and characterization of functional fusion proteins having a collagen-binding domain
Collagen-binding growth factors: Production and characterization of functional fusion proteins having a collagen-binding domain
复制标题
DOI：
10.1073/pnas.95.12.7018
复制
发表时间：
1998-06-09
影响因子：
11.1
通讯作者：
Wada, F
中科院分区：
文献类型：
--
作者：
Nishi, N;Matsushita, O;Wada, F
自分泌/旁分泌肽类信号分子如生长因子等具有多种生物学活性，具有广泛的临床应用前景。然而，人们不能期望通过普通药物递送系统施用的因子的特定治疗效果，因为它们具有有限的靶特异性和短的体内半衰期。为了克服使用生长因子作为治疗剂的困难，我们已经产生了由生长因子部分和来自溶组织梭菌胶原酶的胶原结合结构域（CBD）组成的融合蛋白。在CBD的N端携带表皮生长因子（EGF）或碱性成纤维细胞生长因子（bFGF）的融合蛋白CBEGF/CBFGF与不溶性胶原紧密结合，刺激BALB/c 3 T3成纤维细胞生长;当皮下注射到裸鼠中时，CBEGF在注射部位保持长达10天，而EGF在注射后24小时未检测到。虽然CBEGF在体内没有发挥促生长作用，但CBFGF在注射后5天和7天强烈刺激基质细胞的DNA合成，而不是bFGF。这些结果表明，CBD可以用作锚定单元，以产生在体内不扩散和持久的融合蛋白。
0 The autocrine/paracrine peptide signaling molecules such as growth factors have many promising biologic activities for clinical applications. However, one cannot expect specific therapeutic effects of the factors administered by ordinary drug delivery systems as they have limited target specificity and short half-lives in vivo. To overcome the difficulties in using growth factors as therapeutic agents, we have produced fusion proteins consisting of growth factor moieties and a collagen-binding domain (CBD) derived from Clostridium histolyticum collagenase. The fusion proteins carrying the epidermal growth factor (EGF) or basic fibroblast growth factor (bFGF) at the N terminal of CBD (CBEGF/CBFGF) tightly bound to insoluble collagen and stimulated the growth of BALB/c 3T3 fibroblasts as much as the unfused counterparts; CBEGF, when injected subcutaneously into nude mice, remained at the sites of injection for up to 10 days, whereas EGF was not detectable 24 h after injection. Although CBEGF did not exert a growth-promoting effect in vivo, CBFGF, but not bFGF, strongly stimulated the DNA synthesis in stromal cells at 5 days and 7 days after injection. These results indicate that CBD may be used as an anchoring unit to produce fusion proteins nondiffusible and long-lasting in vivo.