结核分枝杆菌丝氨酸/苏氨酸蛋白激酶PknB的表达与特性分析
Expression and characterization of the Mycobacterium tuberculosis serine/threonine protein kinase PknB.
作者信息
Av-Gay Y, Jamil S, Drews S J
机构信息
Department of Medicine, Division of Infectious Diseases, University of British Columbia, Vancouver, British Columbia, Canada V5Z 3J5.
出版信息
Infect Immun. 1999 Nov;67(11):5676-82. doi: 10.1128/IAI.67.11.5676-5682.1999.
Abstract
PknB is a member of the newly discovered eukaryotic-like protein serine/threonine kinase (PSTK) family of proteins. The pknB gene was cloned and expressed in Escherichia coli. The active recombinant protein was purified and shown to be reactive with antiphosphoserine antibodies, as well as with antibodies to the phosphorylated eukaryotic Ser/Thr kinases mitogen-activated protein kinase kinase 3 and 6, P38, and Creb. In vitro kinase assays demonstrated that PknB is a functional kinase that is autophosphorylated on serine/threonine residues and is also able to phosphorylate the peptide substrate myelin basic protein. Analysis of pknB expression in Mycobacterium tuberculosis indicates the presence of pknB mRNA in (i) organisms grown in vitro in bacteriological media, (ii) a murine macrophage in vitro infection model, and (iii) in vivo alveolar macrophages from a patient with tuberculosis.
摘要
PknB是新发现的类真核蛋白丝氨酸/苏氨酸激酶（PSTK）家族的成员。pknB基因在大肠杆菌中被克隆并表达。活性重组蛋白被纯化，并显示与抗磷酸丝氨酸抗体以及与磷酸化的真核丝氨酸/苏氨酸激酶丝裂原活化蛋白激酶激酶3和6、P38和Creb的抗体有反应。体外激酶分析表明，PknB是一种功能性激酶，它在丝氨酸/苏氨酸残基上自磷酸化，并且还能够磷酸化肽底物髓鞘碱性蛋白。对结核分枝杆菌中pknB表达的分析表明，在以下情况中存在pknB mRNA：（i）在细菌学培养基中体外培养的生物体，（ii）体外小鼠巨噬细胞感染模型，以及（iii）来自结核病患者的体内肺泡巨噬细胞。
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