BARLEY ALEURONE LAYERS SECRETE A NUCLEASE IN RESPONSE TO GIBBERELLIC-ACID - PURIFICATION AND PARTIAL CHARACTERIZATION OF THE ASSOCIATED RIBONUCLEASE, DEOXYRIBONUCLEASE, AND 3'-NUCLEOTIDASE ACTIVITIES
BARLEY ALEURONE LAYERS SECRETE A NUCLEASE IN RESPONSE TO GIBBERELLIC-ACID - PURIFICATION AND PARTIAL CHARACTERIZATION OF THE ASSOCIATED RIBONUCLEASE, DEOXYRIBONUCLEASE, AND 3'-NUCLEOTIDASE ACTIVITIES
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DOI：
10.1104/pp.82.3.801
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发表时间：
1986-11-01
期刊：
影响因子：
7.4
通讯作者：
HO, THD
中科院分区：
文献类型：
--
作者：
BROWN, PH;HO, THD
用赤霉酸孵育大麦（Hordeum vulgare L. cv. Himalaya）半种子增强了糊粉组织中核糖核酸酶和脱氧核糖核酸酶的分泌（MJ Chrispeels，JE Varner 1967 Plant Physiol 42：398-406；L Taiz，JE Starks 1977 Plant Physiol 60： 182-189）。在用赤霉酸孵育的半种子培养基中，这些活性比对照培养基高 50 倍以上。核糖核酸酶和脱氧核糖核酸酶活性最初在激素诱导后24至48小时出现在培养基中，并增加长达96小时。两种活性的最适 pI 为 6.0，最适温度为 55°C。当通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳分析来自赤霉酸处理的半种子的培养基时，主要核糖核酸酶和脱氧核糖核酸酶活性条带发生了共迁移。在采用硫酸铵分级分离的 2,700 倍纯化过程中，两种酶的活性始终保持相关。肝素-琼脂糖亲和层析和活性蓝2-琼脂糖亲和层析。在整个纯化过程中，伴随着核糖核酸酶和脱氧核糖核酸酶活性的是水解3''-AMP的3''-磷酸酯键的能力。纯化的蛋白质由单一多肽组成，经十二烷基硫酸钠聚丙烯酰胺凝胶电泳测定，表观分子量为36千道尔顿。结论是，响应赤霉酸，大麦糊粉组织分泌具有核糖核酸酶、脱氧核糖核酸酶和3''-核苷酸酶活性的核酸酶。
Incubation of barley (Hordeum vulgare L. cv. Himalaya) half-seeds with gibberellic acid enhances the secretion of ribonuclease and deoxyribonuclease from aleurone tissue (MJ Chrispeels, JE Varner 1967 Plant Physiol 42: 398-406; L Taiz, JE Starks 1977 Plant Physiol 60: 182-189). These activities were over 50-fold greater in medium of half-seeds incubated with gibberellic acid than in control medium. Ribonuclease and deoxyribonuclease activities initially appeared in the medium 24 to 48 hours after hormone induction and increased for up to 96 hours. Both activities had a pI optimum of 6.0 and a temperature optimum of 55.degree.C. When the medium from gibberellic acid-treated half-seeds was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis, the major ribonuclease and deoxyribonuclease activity bands comigrated. The two enzyme activities remained associated throughout a 2,700-fold purification employing ammonium sulfate fractionation. Heparin-Agarose affinity chromatography, and Reactive Blue 2-Agarose affinity chromatography. Also accompanying the ribonucelase and deoxyribonuclease activities throughout purification was the ability to hydrolyze the 3''-phosphoester linkage of 3''-AMP. The purified protein was composed of a single polypeptide with an apparent molecular weight of 36 kilodaltons as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. It is concluded that in response to gibberellic acid, barley aleurone tissue secretes a nuclease having ribonuclease, deoxyribonuclease, and 3''-nucleotidase activities.