Open AccessMass spectral determination of phosphopantetheinylation specificity for carrier proteins in Mycobacterium tuberculosis
Author(s) -
Jung James,
Bashiri Ghader,
Johnston Jodie M.,
Baker Edward N.
Publication year - 2016
Publication title -
febs open bio
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.718
H-Index - 31
ISSN - 2211-5463
DOI - 10.1002/2211-5463.12140
Subject(s) - polyketide , mycobacterium tuberculosis , serine , biochemistry , acyl carrier protein , chemistry , identification (biology) , biology , mycobacterium , computational biology , bacteria , tandem mass spectrometry , biosynthesis , tuberculosis , mass spectrometry , enzyme , genetics , chromatography , medicine , pathology , botany
Phosphopantetheinyl transferases ( PPT ases) are key elements in the modular syntheses performed by multienzyme systems such as polyketide synthases. PPT ases transfer phosphopantetheine derivatives from Coenzyme A to carrier proteins ( CP s), thus orchestrating substrate supply. We describe an efficient mass spectrometry‐based protocol for determining CP specificity for a particular PPT ase in organisms possessing several candidate PPT ases. We show that the CP s MbtL and PpsC, both involved in synthesis of essential metabolites in Mycobacterium tuberculosis , are exclusively activated by the type 2 PPT ase PptT and not the type 1 AcpS. The assay also enables conclusive identification of the reactive serine on each CP .