Vassimon C S, Gomes E T, Rothschild A M
Agents Actions. 1987 Jun;21(1-2):62-5. doi: 10.1007/BF01974922.
Rat peritoneal fluid mast cell present parallel increases in cell area (swelling), and in hydrolytic activity on the trypsin substrate p-tosyl arginine methyl ester (TAME), when placed in Tris buffers of concentrations between 0.15 and 0.03 M. Under these conditions, cells do not degranulate and preserve their trypsin-like enzyme activity after low speed centrifugation. Exposure to more dilute Tris buffers, between 0.015 and 0.003 M, leads to cell rupture accompanied by progressive degranulation and loss of activity on TAME. Protamine, a heparin antagonist prevented this loss when added to mast cells prior to hyposmotic lysis, or lysis by sonication or repeated periods of freezing and thawing. Enzyme activity released in the presence of protamine was fully recovered in supernates of cell lysates submitted to low speed centrifugation. Controlled swelling of mast cells propitiates the expression of trypsin-like activity, possibly by facilitating enzyme-substrate interaction. Cell lysis on the contrary, leads to inactivation of such activity, possibly by enzyme binding to heparin in exposed mast cell granules.
当置于浓度在0.15至0.03 M之间的Tris缓冲液中时,大鼠腹膜液肥大细胞的细胞面积(肿胀)以及对胰蛋白酶底物对甲苯磺酰精氨酸甲酯(TAME)的水解活性会同时增加。在这些条件下,细胞不会脱颗粒,并且在低速离心后仍保留其类胰蛋白酶活性。暴露于浓度在0.015至0.003 M之间的更稀Tris缓冲液中会导致细胞破裂,并伴有逐渐脱颗粒以及对TAME活性的丧失。鱼精蛋白是一种肝素拮抗剂,在低渗裂解、超声裂解或反复冻融之前添加到肥大细胞中时,可防止这种活性丧失。在鱼精蛋白存在下释放的酶活性在经过低速离心的细胞裂解物上清液中可完全恢复。肥大细胞的可控肿胀可能通过促进酶 - 底物相互作用来促进类胰蛋白酶活性的表达。相反,细胞裂解会导致这种活性失活,可能是由于酶与暴露的肥大细胞颗粒中的肝素结合。