Jung James, Bashiri Ghader, Johnston Jodie M, Baker Edward N
Maurice Wilkins Centre for Molecular Biodiscovery and School of Biological Sciences The University of Auckland New Zealand; Present address: W. M. Keck Structural Biology Laboratory Cold Spring Harbor Laboratory 1 Bungtown Road Cold Spring Harbor NY 11724 USA.
Maurice Wilkins Centre for Molecular Biodiscovery and School of Biological Sciences The University of Auckland New Zealand.
FEBS Open Bio. 2016 Oct 24;6(12):1220-1226. doi: 10.1002/2211-5463.12140. eCollection 2016 Dec.
Phosphopantetheinyl transferases (PPTases) are key elements in the modular syntheses performed by multienzyme systems such as polyketide synthases. PPTases transfer phosphopantetheine derivatives from Coenzyme A to carrier proteins (CPs), thus orchestrating substrate supply. We describe an efficient mass spectrometry-based protocol for determining CP specificity for a particular PPTase in organisms possessing several candidate PPTases. We show that the CPs MbtL and PpsC, both involved in synthesis of essential metabolites in , are exclusively activated by the type 2 PPTase PptT and not the type 1 AcpS. The assay also enables conclusive identification of the reactive serine on each CP.
磷酸泛酰巯基乙胺基转移酶（PPTases）是多酶系统（如聚酮合酶）进行模块化合成的关键要素。PPTases将磷酸泛酰巯基乙胺衍生物从辅酶A转移到载体蛋白（CPs）上，从而协调底物供应。我们描述了一种基于质谱的高效方法，用于确定具有多种候选PPTases的生物体中特定PPTase的CP特异性。我们发现，参与[具体生物体]必需代谢物合成的CPs MbtL和PpsC仅由2型PPTase PptT激活，而非1型AcpS。该检测方法还能够确凿地鉴定每个CP上的反应性丝氨酸。
