Lian Z, Tianjue Y
Department of New Antibiotics, Institute of Medicinal Biotechnology, CAMS &. PUMC, Beijing 100050.
Chin Med Sci J. 1999 Mar;14(1):57-60.
The present study identified the properties of porins in the outer membrane in Pseudomonas aeruginosa, and showed the role of outer membrane in determining imipenem diffusion in Pseudomonas aeruginosa. The molecular weight of the major outer membrane protein was analyzed by SDS-PAGE. The purification of the porins in Pseudomonas aeruginosa was achieved by DEAE ion-exchange HPLC. The purified outer membrane proteins were reconstituted with phosphatidylcholine and dicetylphosphate into membrane vesicles, and were tested by the liposomes swelling method for the diffusion of imipenem. The permeability assay showed that OprC (70 kD), OprD2 (46 kD), and OprE (43 kD) were the channel-forming proteins. But only OprD2 was thought to be the likely route of imipenem diffusion.
本研究确定了铜绿假单胞菌外膜中孔蛋白的特性，并展示了外膜在决定亚胺培南在铜绿假单胞菌中扩散的作用。通过SDS-PAGE分析主要外膜蛋白的分子量。采用DEAE离子交换HPLC法实现了铜绿假单胞菌中孔蛋白的纯化。将纯化的外膜蛋白与磷脂酰胆碱和磷酸二鲸蜡酯重构为膜囊泡，并通过脂质体肿胀法检测亚胺培南的扩散情况。通透性测定表明，OprC（70 kD）、OprD2（46 kD）和OprE（43 kD）是形成通道的蛋白。但只有OprD2被认为可能是亚胺培南扩散的途径。
