Brown M S, Segal A, Stadtman E R
Proc Natl Acad Sci U S A. 1971 Dec;68(12):2949-53. doi: 10.1073/pnas.68.12.2949.
Earlier studies showed that two protein components, P(I) and P(II), are concerned with the adenylylation and deadenylylation of Escherichia coli glutamine synthetase (EC 6.3.1.2). P(I) by itself catalyzes both adenylylation and deadenylylation, but its activity is modulated by the P(II)-protein and by glutamine, 2-oxoglutarate, ATP, and UTP, The P(II)-protein exists in two forms: one form, P(II)-AT, stimulates P(I)-catalyzed adenylylation activity in the absence of glutamine and makes this activity very sensitive to inhibition by 2-oxoglutarate; it does not affect deadenylylation activity. The other form, P(II)-DA, stimulates adenylylation only if glutamine is present, and also stimulates the deadenylylation activity of P(I), which is then dependent upon the presence of ATP and 2-oxoglutarate. Conversion of P(II)-AT to P(II)-DA requires the presence of UTP, ATP, and 2-oxoglutarate; it is catalyzed by an enzyme present in P(I) preparations. UTP may be directly involved in this conversion since P(II)-DA fractions reisolated by filtration through Sephadex G-100 contain small quantities of a bound uridine derivative that lacks the gamma-phosphoryl group of UTP. The activity of P(II)-DA, but not of P(II)-AT, is destroyed by treatment with snake-venom phosphodiesterase. ATP and 2-oxoglutarate apparently function as allosteric effectors for the conversion of P(II)-AT to P(I)-DA.
早期研究表明,两种蛋白质成分P(I)和P(II)与大肠杆菌谷氨酰胺合成酶(EC 6.3.1.2)的腺苷酰化和去腺苷酰化有关。P(I)本身催化腺苷酰化和去腺苷酰化,但它的活性受P(II)蛋白以及谷氨酰胺、2-氧代戊二酸、ATP和UTP的调节。P(II)蛋白以两种形式存在:一种形式是P(II)-AT,在没有谷氨酰胺的情况下刺激P(I)催化的腺苷酰化活性,并使该活性对2-氧代戊二酸的抑制非常敏感;它不影响去腺苷酰化活性。另一种形式是P(II)-DA,仅在存在谷氨酰胺时刺激腺苷酰化,并且还刺激P(I)的去腺苷酰化活性,此时该活性依赖于ATP和2-氧代戊二酸的存在。P(II)-AT向P(II)-DA的转化需要UTP、ATP和2-氧代戊二酸的存在;它由P(I)制剂中存在的一种酶催化。UTP可能直接参与这种转化,因为通过Sephadex G-100过滤重新分离的P(II)-DA级分含有少量结合的尿苷衍生物,该衍生物缺乏UTP的γ-磷酸基团。用蛇毒磷酸二酯酶处理会破坏P(II)-DA的活性,但不会破坏P(II)-AT的活性。ATP和2-氧代戊二酸显然作为变构效应剂参与P(II)-AT向P(I)-DA的转化。