PHARMACOLOGICAL ACTIVITY OF THE C-TERMINAL AND N-TERMINAL DOMAINS OF SECRETORY LEUKOPROTEASE INHIBITOR IN-VITRO
PHARMACOLOGICAL ACTIVITY OF THE C-TERMINAL AND N-TERMINAL DOMAINS OF SECRETORY LEUKOPROTEASE INHIBITOR IN-VITRO
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DOI：
10.1111/j.1476-5381.1995.tb15892.x
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发表时间：
1995-07-01
影响因子：
7.3
通讯作者：
SUZUKI, Y
中科院分区：
文献类型：
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作者：
MASUDA, K;KAMIMURA, T;SUZUKI, Y
1为了研究分泌型白细胞蛋白酶抑制剂（SLPI）结构域的生理功能，研究了半长度SLPI（Ser 1-Pro54）SLPI和（Asn 55-Ala 107）SLPI的生物学活性，并与全长SLPI的生物学活性进行了比较。C-末端结构域（Asn 55-Ala 107）SLPI对人中性粒细胞弹性蛋白酶（NE）、组织蛋白酶G和糜蛋白酶的抑制能力与全长SLPI一样强。其胰蛋白酶抑制活性低于完整的SLPI。对于SLPI的N-末端结构域，（Ser 1-Pro54）SLPI，在本研究中未检测到针对测试的丝氨酸蛋白酶的抑制活性。（Asn 55-Ala 107）SLPI对NE对天然底物弹性蛋白和胶原蛋白的蛋白水解的抑制作用与全SLPI相当（弹性蛋白，对于SLPI，IC 50 = 907 +/- 31 nM，对于（Asn 55-Ala 107）SLPI，IC 50 = 767 +/- 33 nM;胶原蛋白，SLPI的IC 50 = 862 +/- 36 nM，727 +/- 47 nM（Asn 55-Ala 107）SLPI）。4通过亲和柱色谱法测量全长和半长SLPI与肝素的结合亲和力。全长SLPI显示出对肝素的高亲和力，而两种半长SLPI的结合能力较低。（用于洗脱的NaCl浓度，SLPI为0.45 M，（Ser 1-Pro54）SLPI为0.24 M，（Asn 55-Ala 107）SLPI为0.27 M）。5使用活化部分凝血活酶时间（APTT）测量全SLPI和（Asn 55-Ala 107）SLPI对血液凝固的影响。全长SLPI剂量依赖性地延长凝血时间（1.25、2.5和5.0 μ M），而（Asn 55-Ala 107）SLPI即使在最高浓度下也没有效果。6总之，SLPI的C-末端结构域是治疗炎性疾病的有希望的候选者，其中嗜中性粒细胞蛋白酶的参与已被提出。
1 In order to characterize the physiological functions of the domain structure of secretory leukoprotease inhibitor (SLPI), the biological capacities of half-length SLPIs, (Ser1-Pro54)SLPI and (Asn55-Ala107)SLPI, were investigated and compared with those of full-length SLPI.2 The activities of these inhibitors against several serine proteases were determined using synthetic chromogenic substrates. The inhibitory capacity of the C-terminal domain, (Asn55-Ala107)SLPI, was as strong as that of full-length SLPI against human neutrophil elastase (NE), cathepsin G and chymotrypsin. It possessed less trypsin inhibitory activity than intact SLPI. For the N-terminal domain of SLPI, (Ser1-Pro54)SLPI, no inhibitory activity could be detected against the serine proteases tested in this study.3 The inhibitory activity of (Asn55-Ala107)SLPI against the proteolysis of the natural substrates elastin and collagen by NE was comparable with that of full-SLPI (elastin, IC50 = 907 +/- 31 nM for SLPI, 767 +/- 33 nM for (Asn55-Ala107)SLPI; collagen, IC50 = 862 +/- 36 nM for SLPI, 727 +/- 47 nM for (Asn55-Ala107)SLPI).4 The binding affinities of full- and half-length SLPIs for heparin were measured by affinity column chromatography. Full-length SLPI showed high affinity for heparin while the binding capacities of both half-length SLPIs were lower. (Concentration of NaCl for elution, 0.45 M for SLPI, 0.24 M for (Ser1-Pro54)SLPI, 0.27 M for (Asn55-Ala107)SLPI).5 The effects of full-SLPI and (Asn55-Ala107)SLPI on blood coagulation were measured using the activated partial thromboplastin time (APTT). Full-length SLPI prolonged clotting time dose-dependently (1.25, 2.5 and 5.0 mu M), whereas (Asn55-Ala107)SLPI had no effect even at the highest concentration.6 In conclusion, the C-terminal domain of SLPI is a promising candidate for the treatment of inflammatory diseases in which participation of neutrophil proteases has been suggested.