The reversible dissociation of the alkaline phosphatase of Escherichia coli. II. Properties of the subunit.
The reversible dissociation of the alkaline phosphatase of Escherichia coli. II. Properties of the subunit.
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大肠杆菌碱性磷酸酶的可逆解离。
DOI：
10.1016/s0021-9258(18)97058-4
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发表时间：
1965
期刊：
影响因子：
--
通讯作者：
M. Schlesinger
中科院分区：
文献类型：
--
作者：
M. Schlesinger
方法酶和测定法--高纯度碱性磷酸酶的制备和酶活性测定方法与前文相同。如Byrne（7）所述制备14 C-精氨酸标记的酶。将与抗体-酶和兔抗血清的反应物在总体积为1.0 ml的条件下于4”孵育16小时。离心收集沉淀，用冷NaCl洗涤三次，再用1.0ml 0.5%十二烷基硫酸钠溶解，在278 mp下测定吸光度。测定上清液部分和再溶解的沉淀物的等分试样的酶活性。将在1.0 ml中制备的亚基与高碘酸钠（0.003 M）在pH 2.0、pH 2.0和pH 2.0下孵育，
MethodsEnzyme and Assays-Preparations of highly purified alkaiine phosphatase and the procedure for assay of enzymic activity are identical with those described in the preceding paper. Enzyme labeled with 14C-arginine was prepared as described by Byrne (7). Reaction with Antibody-Enzyme and rabbit antiserum were incubated at 4” for 16 hours in a total volume of 1.0 ml. The precipitates were collected by centrifugation, washed three times with cold NaCl, and redissolved with 1.0 ml of 0.5% sodium dodecyl sulfate, and the absorbance was measured at 278 mp. Aliquots of the supernatant fractions and the redissolved precipitates were assayed for enzymic activity. Reaction with Period&e-Acid-prepared subunits in 1.0 ml were incubated with sodium periodate (0.003 M) at either pH 2.0, pH