HEMOLYTIC-ACTIVITY IN THE PERIODONTOPATHOGEN PORPHYROMONAS-GINGIVALIS - KINETICS OF ENZYME-RELEASE AND LOCALIZATION
HEMOLYTIC-ACTIVITY IN THE PERIODONTOPATHOGEN PORPHYROMONAS-GINGIVALIS - KINETICS OF ENZYME-RELEASE AND LOCALIZATION
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DOI：
10.1128/iai.59.6.1932-1940.1991
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发表时间：
1991-06-01
影响因子：
3.1
通讯作者：
HOLT, SC
中科院分区：
文献类型：
--
作者：
CHU, L;BRAMANTI, TE;HOLT, SC
对牙龈卟啉单胞菌W50、W83、A7A1 - 28和ATCC 33277裂解绵羊、人类和兔子红细胞的能力进行了研究。所研究的所有牙龈卟啉单胞菌菌株在生长过程中都产生了活跃的溶血活性，在指数生长后期 - 稳定生长前期活性最强。该酶与细胞结合且与外膜相关。对牙龈卟啉单胞菌W50进行分级分离后发现，假定的溶血素几乎完全定位在外膜组分中，显著的溶血活性集中在外膜囊泡中。Ca²⁺和Mg²⁺离子显著提高了溶血活性的表达。溶血活性被蛋白酶K、胰蛋白酶、蛋白酶抑制剂对甲苯磺酰 - L - 赖氨酸氯甲基酮和苄脒、代谢抑制剂间氯苯腙以及碘乙酸抑制。氰化钾和叠氮化钠（NaN₃）仅部分抑制牙龈卟啉单胞菌的溶血活性，而针对每种牙龈卟啉单胞菌菌株全细胞的抗血清对溶血活性有显著的抑制作用。牙龈卟啉单胞菌W50溶血素在至少10 mM的浓度下被半胱氨酸、二硫苏糖醇和谷胱甘肽抑制；在低浓度（即2 mM）下，二硫苏糖醇不能完全抑制溶血活性。加热到55℃以上会导致溶血活性几乎完全被抑制。血红素限制（即铁）对溶血素产生的影响表明，与在过量血红素存在下生长的牙龈卟啉单胞菌相比，血红素的限制或缺乏会导致溶血素产生显著增加。
Porphyromonas gingivalis W50, W83, A7A1-28, and ATCC 33277 were investigated for their abilities to lyse sheep, human, and rabbit erythrocytes. All of the P. gingivalis strains studied produced an active hemolytic activity during growth, with maximum activity occurring in late-exponential-early-stationary growth phase. The enzyme was cell bound and associated with the outer membrane. Fractionation of P. gingivalis W50 localized the putative hemolysin almost exclusively in the outer membrane fraction, with significant hemolytic activity concentrated in the outer membrane vesicles. Ca2+ and Mg2+ ions significantly increased the expression of hemolytic activity. Hemolytic activity was inhibited by proteinase K, trypsin, the proteinase inhibitors Na-P-tosyl-L-lysine chloromethyl ketone and benzamidine, the metabolic inhibitor M-chlorophenyl-hydrazone, and iodoacetate. KCN and sodium azide (NaN3) only partially inhibited P. gingivalis hemolytic activity, while antiserum to whole cells of each of the P. gingivalis strains had a significant inhibitory effect on hemolytic activity. The P. gingivalis W50 hemolysin was inhibited by cysteine, dithiothreitol, and glutathione at concentrations of at least 10 mM; at low concentrations (i.e., 2 mM), dithiothreitol did not completely inhibit hemolytic activity. Heating to temperatures above 55-degrees-C resulted in an almost complete inhibition of hemolytic activity. The effect of heme limitation (i.e., iron) on hemolysin production indicated that either limitation or starvation for heme resulted in significantly increased hemolysin production compared with that of P. gingivalis grown in the presence of excess heme.