. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. UML'LUS BLOOD CELL SECRETES «rMACROGLOBULIN 141 E c O) o (O - 20 40 Minutes 60 80 1 00 Figure 3. Demonstration of a:-macroglobulin in materials released by saline-washed Limuliis blood cells stimulated to degranulate by ex- posure to the ionophore, A23187, and in plasma from the same animal. The released materials protect trypsin from inactivation by the high molecular mass active site inhibitor, soybean trypsin inhibitor. Ten ^g samples of trypsin were preincubated with 20 M' of plasma (curve B), the
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. UML'LUS BLOOD CELL SECRETES «rMACROGLOBULIN 141 E c O) o (O - 20 40 Minutes 60 80 1 00 Figure 3. Demonstration of a:-macroglobulin in materials released by saline-washed Limuliis blood cells stimulated to degranulate by ex- posure to the ionophore, A23187, and in plasma from the same animal. The released materials protect trypsin from inactivation by the high molecular mass active site inhibitor, soybean trypsin inhibitor. Ten ^g samples of trypsin were preincubated with 20 M' of plasma (curve B), the materials released from 12 ^1 of packed cells, which was the amount of cells contained in 530 /jl of whole blood (curve C). or 160 ^1 of the penultimate saline wash (curve D) for 10 min and then 20 ^g of soybean trypsin inhibitor was added. The remaining active trypsin ( the trypsin bound to the «2-macroglobulin in the sample, and thereby pro- tected from inactivation by soybean trypsin inhibitor) was assayed by its ability to hydrolyze the low molecular mass amide substrate, BAPNA. This was followed by the increase in optical absorbance at 410 nm. In the absence of added «:-macroglobulin, the hydrolysis of BAPNA is zero (not shown). Curve A is the activity of 10 /ig of trypsin in the absence of soybean trypsin inhibitor or «:-macroglobulin. In this sample, the cells from a given volume of blood (curve C) contained as much a^-macroglobulin as the plasma from the same volume of blood (curve B). In other trials, the cells have contained as much as 2% of the total «:-macroglobulin in a given volume of blood. the cells and then their lysis in distilled water. The effi- ciency of washing is indicated by the minimal contami- nation of the LAL preparations with hemocyanin (com- pare the relative intensities of the hemocyanin band at 67 kDa and the «rmacroglobulin band at 185 kDa in plasma (Fig. 2, lanes and ) with that in LAL (Fig. 4, lanes and ). We spe
Size: 1585px × 1575px
Photo credit: © Library Book Collection / Alamy / Afripics
License: Licensed
Model Released: No
Keywords: ., bookauthorlilliefrankrat, booksubjectbiology, booksubjectzoology
