AZUROCIDIN AND A HOMOLOGOUS SERINE PROTEASE FROM NEUTROPHILS - DIFFERENTIAL ANTIMICROBIAL AND PROTEOLYTIC PROPERTIES
AZUROCIDIN AND A HOMOLOGOUS SERINE PROTEASE FROM NEUTROPHILS - DIFFERENTIAL ANTIMICROBIAL AND PROTEOLYTIC PROPERTIES
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DOI：
10.1172/jci114518
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发表时间：
1990-03-01
影响因子：
15.9
通讯作者：
GABAY, JE
中科院分区：
文献类型：
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作者：
CAMPANELLI, D;DETMERS, PA;GABAY, JE
通过对azurophil颗粒膜相关物质的酸萃取，凝胶过滤和反相层析，从人中性粒细胞中纯化出两个29-kD多肽azuroidin和p29b。azuroidin和p29b具有nh2末端序列同源性，与弹性蛋白酶、组织蛋白酶G等丝氨酸蛋白酶具有同源性，p29b结合[3H]氟磷酸二异丙基和水解弹性蛋白、酪蛋白和血红蛋白。p29b的肽底物无法确定。唑虫啶既没有结合[3H]氟磷酸二异丙基，也没有水解任何被测试的蛋白质、肽或酯。在杀微生物试验中，纯化的灭虫脒对大肠杆菌、粪链球菌和白色念珠菌的活性与p29b相当。抑菌活性在轻度酸性条件下增强，但被NaCl、CaCl2和血清呈剂量依赖性抑制。免疫印迹分析用单特异性抗体定位> 90%的azuroidin和> 75%的p29b到PMN裂解物富含azurophil颗粒的部分。免疫电镜证实了偶氮菌素定位于偶氮粒细胞颗粒。偶氮胞嘧啶与偶氮颗粒膜相关，但不是一个完整的膜蛋白。因此，azuroidin和p29b是储存在azurophil颗粒中的丝氨酸蛋白酶同源物家族的成员，并可能在涉及PMN的炎症和抗菌过程中发挥作用。
Two 29-kD polypeptides, azurocidin and p29b, were purified to homogeneity from human neutrophils by acid extraction of azurophil granule membrane-associated material followed by gel filtration and reverse-phase chromatography. Azurocidin and p29b share NH2-terminal sequence homology with each other as well as with elastase, cathepsin G, and other serine proteases, p29b bound [3H]diisopropyl fluorophosphate and hydrolyzed elastin, casein, and hemoglobin. A peptide substrate for p29b could not be identified. Azurocidin neither bound [3H]diisopropyl fluorophosphate nor hydrolyzed any of the proteins, peptides, or esters tested. In microbicidal assays, purified azurocidin was comparable to p29b in activity against Escherichia coli, Streptococcus faecalis, and Candida albicans. The antimicrobial activity of azurocidin was enhanced under mildly acidic conditions, but was inhibited in a dose-dependent manner by NaCl, CaCl2, or serum. Immunoblot analysis with monospecific antibodies localized > 90% of the azurocidin and > 75% of the p29b to azurophil granule-rich fractions of PMN lysates. Immunoelectron microscopy confirmed the localization of azurocidin to the azurophil granules. Azurocidin associated with the azurophil granule membrane, but did not appear to be an integral membrane protein. Thus, azurocidin and p29b are members of a family of serine protease homologs stored in azurophil granules and may play a role in inflammatory and antimicrobial processes involving PMN.