Noseworthy J, Smith G H, Himmelhoch S R, Evans W H
J Cell Biol. 1975 Jun;65(3):577-86. doi: 10.1083/jcb.65.3.577.
The postnuclear supernatant fraction of sucrose homogenates of guinea pig polymorphonuclear leukocytes (PMNL) was subjected to differential centrifugation to obtain a total particulate fraction, a particle-free supernatant fraction, highly enriched fractions of primary and secondary granules, and a membrane-rich fraction. The various fractions were solubilized in buffer containing sodium dodecyl sulfate (SDS) and analyzed for protein and glycoproteincomponents by SDS -polyacrylamide gel electrophoresis. The major glycoprotein components of the postnuclear supernatant fraction were found mainly associated with the enriched fraction of secondary granules and, to a lesser extent, with the membrane-rich fraction. No major glycoprotein components were visible in the polypeptide electrophoretic patterns of the primary granule fraction or of the particle-free supernate. Attempts at separation of guinea pig granules by zonal sucrose density gradient centrifugation were only partially successful. Data supporting a species difference in this regard between rabbit and guinea pig PMNL granules are presented.
对豚鼠多形核白细胞(PMNL)蔗糖匀浆的核后上清液部分进行差速离心,以获得总颗粒部分、无颗粒上清液部分、高度富集的初级和次级颗粒部分以及富含膜的部分。将各部分溶解于含有十二烷基硫酸钠(SDS)的缓冲液中,并通过SDS-聚丙烯酰胺凝胶电泳分析蛋白质和糖蛋白成分。发现核后上清液部分的主要糖蛋白成分主要与次级颗粒的富集部分相关,在较小程度上与富含膜的部分相关。在初级颗粒部分或无颗粒上清液的多肽电泳图谱中未可见主要糖蛋白成分。通过区带蔗糖密度梯度离心分离豚鼠颗粒的尝试仅部分成功。本文提供了支持兔和豚鼠PMNL颗粒在这方面存在种属差异的数据。