LAMELLAR BODY-ENRICHED FRACTIONS FROM NEONATAL MICE - PREPARATIVE TECHNIQUES AND PARTIAL CHARACTERIZATION
LAMELLAR BODY-ENRICHED FRACTIONS FROM NEONATAL MICE - PREPARATIVE TECHNIQUES AND PARTIAL CHARACTERIZATION
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DOI：
10.1111/1523-1747.ep12276826
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发表时间：
1985-01-01
影响因子：
6.5
通讯作者：
ELIAS, PM
中科院分区：
文献类型：
--
作者：
GRAYSON, S;JOHNSONWINEGAR, AG;ELIAS, PM
有几个问题阻碍了从哺乳动物表皮中分离板层小体。我们通过利用葡萄球菌表皮溶解毒素提供完整的外表皮片，通过在细胞破碎器中受控的均质，并通过匀浆通过一系列分级的核孔过滤器，获得了富含完整LB的颗粒(Science 221：962,1983)。这种制剂比各种不同或蔗糖/甲基咪唑胺不连续离心法制备的组分含有更多完整的Lb。对该组分的酶含量初步鉴定表明，它富含某些水解酶(酸性磷酸酶、羧基肽酶、组织蛋白酶B、酸性脂肪酶、鞘磷脂酶和磷脂酶A)，但显著缺乏所有的硫酸酯酶、β-葡萄糖醛酸酶和非溶酶体蛋白水解酶，即纤溶酶原激活剂。因此，尽管某些特有的溶酶体酶明显缺失，但Lb仍显示出一些溶酶体的特性。板层体组分的单位重量含脂量是匀浆的2-3倍，富含磷脂、游离甾醇和鞘糖脂，但不含其他中性脂或神经酰胺。综上所述，虽然LB中的一些酶可以参与LB脂前体向疏水屏障成分的代谢，但其他酶可能攻击细胞间成分，最终导致脱屑。这些细胞器的脂质分布表明，它们将通透性屏障脂质的前体输送到细胞间域。
Several problems have frustrated the isolation of lamellar bodies (LB) from mammalian epidermis. We obtained pellets enriched in intact LB by utilizing the staphylococcal epidermolytic toxin to provide intact, outer epidermal sheets, by controlled homogenization in a cell disrupter, and by passage of homogenates through a graded series of nucleopore filters (Science 221:962, 1983). Such preparations contained more intact LB than did fractions prepared by a variety of differential or sucrose/metrizamide discontinuous centrifugation methods. Initial characterization of the enzymatic content of this fraction revealed it to be enriched in certain hydrolytic enzymes (acid phosphatase, carboxypeptidase, cathepsin B, acid lipase, sphingomyelinase, and phospholipase A), but strikingly depleted in all sulfatases, .beta.-glucuronidase, and the non-lysosomal protease, plasminogen activator. Thus, LB show some properties of lysosomes, although certain characteristic lysosomal enzymes are strikingly absent. Lamellar body fractions contained 2-3 times more lipid per unit weight than did homogenates, and were enriched in phospholipids, free sterols, and glycosphingolipids, but not in other neutral lipids or ceramides. In summary, whereas some of the enzymes in LB could participate in the metabolism of LB lipid precursors to hydrophobic barrier constituents, others may attack intercellular constituents, ultimately resulting in desquamation. The lipid profile of these organelles suggests that they deliver precursors of permeability barrier lipids to intercellular domains.