TY - JOUR
T1 - Biochemical characterization of phosphoserine phosphatase SerB2 from Mycobacterium marinum
AU - Pierson, Elise
AU - Wouters, Johan
N1 - Funding Information:
EP acknowledges the Fonds de la Recherche Scientifique (F.R.S. -FNRS, Belgium) for her Research Fellow grant.
Funding Information:
The authors would like to thank Adrien Lesne for his valuable contribution to the experiments. The pAVA0421-serb2 plasmid was generously provided by the Seattle Structural Genomics Center for Infectious Disease ( www.SSGCID.org ) which is supported by Federal Contract No. HHSN272201700059C from the National Institute of Allergy and Infectious Diseases , National Institutes of Health , Department of Health and Human Services .
Funding Information:
EP acknowledges the Fonds de la Recherche Scientifique (F.R.S. -FNRS, Belgium) for her Research Fellow grant.The authors would like to thank Adrien Lesne for his valuable contribution to the experiments. The pAVA0421-serb2 plasmid was generously provided by the Seattle Structural Genomics Center for Infectious Disease (www.SSGCID.org) which is supported by Federal Contract No. HHSN272201700059C from the National Institute of Allergy and Infectious Diseases, National Institutes of Health, Department of Health and Human Services.
Publisher Copyright:
© 2020 Elsevier Inc.
PY - 2020/10/1
Y1 - 2020/10/1
N2 - SerB2 is an essential phosphoserine phosphatase (PSP) that has been shown to be involved in Mycobacterium tuberculosis (Mtb) immune evasion mechanisms, and a drug target for the development of new antitubercular agents. A highly similar (91.0%) orthologous enzyme exists in the surrogate organism Mycobacterium marinum (Mma) and could have acquired similar properties. By homology modeling, we show that the two PSPs are expected to exhibit almost identical architectures. MmaSerB2 folds into a homodimer formed by two intertwined subunits including two ACT regulatory domains followed by a catalytic core typical of HAD (haloacid dehalogenase) phosphatases. Their in vitro catalytic properties are closely related as MmaSerB2 also depends on Mg2+ for the dephosphorylation of its substrate, O-phospho-l-serine (PS), and is most active at neutral pH and temperatures around 40 °C. Moreover, an enzyme kinetics study revealed that the enzyme is inhibited by PS as well, but at lower concentrations than MtbSerB2. Substrate inhibition could occur through the binding of PS in the second active site and/or at the ACT domains interface. Finally, previously described beta-carboline MtbSerB2 inhibitors also decrease the phosphatase activity of MmaSerB2. Altogether, these results provide useful information when M.marinum is used as a model to study immune evasion in tuberculosis.
AB - SerB2 is an essential phosphoserine phosphatase (PSP) that has been shown to be involved in Mycobacterium tuberculosis (Mtb) immune evasion mechanisms, and a drug target for the development of new antitubercular agents. A highly similar (91.0%) orthologous enzyme exists in the surrogate organism Mycobacterium marinum (Mma) and could have acquired similar properties. By homology modeling, we show that the two PSPs are expected to exhibit almost identical architectures. MmaSerB2 folds into a homodimer formed by two intertwined subunits including two ACT regulatory domains followed by a catalytic core typical of HAD (haloacid dehalogenase) phosphatases. Their in vitro catalytic properties are closely related as MmaSerB2 also depends on Mg2+ for the dephosphorylation of its substrate, O-phospho-l-serine (PS), and is most active at neutral pH and temperatures around 40 °C. Moreover, an enzyme kinetics study revealed that the enzyme is inhibited by PS as well, but at lower concentrations than MtbSerB2. Substrate inhibition could occur through the binding of PS in the second active site and/or at the ACT domains interface. Finally, previously described beta-carboline MtbSerB2 inhibitors also decrease the phosphatase activity of MmaSerB2. Altogether, these results provide useful information when M.marinum is used as a model to study immune evasion in tuberculosis.
KW - Mycobacterium marinum
KW - Phosphoserine phosphatase
KW - HAD phosphatases
KW - clofazimine
KW - Harmine derivatives
KW - Mycobacterium marinum
KW - SerB2
KW - Phosphoserine phosphatase
KW - HAD phosphatases
KW - clofazimine
KW - Harmine derivatives
UR - http://www.scopus.com/inward/record.url?scp=85089185281&partnerID=8YFLogxK
U2 - 10.1016/j.bbrc.2020.07.017
DO - 10.1016/j.bbrc.2020.07.017
M3 - Article
SN - 0006-291X
VL - 530
SP - 739
EP - 744
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 4
ER -