Accepted Manuscript Cytoplasmic Peptidoglycan Intermediate Levels in Staphylococcus aureus Harika Vemula, Navid J. Ayon, William G. Gutheil PII:
S0300-9084(15)00364-8
DOI:
10.1016/j.biochi.2015.11.017
Reference:
BIOCHI 4884
To appear in:
Biochimie
Received Date: 11 May 2015 Accepted Date: 18 November 2015
Please cite this article as: H. Vemula, N.J. Ayon, W.G. Gutheil, Cytoplasmic Peptidoglycan Intermediate Levels in Staphylococcus aureus, Biochimie (2015), doi: 10.1016/j.biochi.2015.11.017. This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
L-Ala MurA/Z
MurB
MurC
MurD
D-Ala
Ddl ATP
MurE
100000
D-Ala-D-Ala
MurF
PEP NADPH ATP ATP ATP ATP UDP-NAG UDP-Tri UDP-Penta UDP-EP UDP-NAM UDP-Mono UDP-Di
10000 1000
PG
µM
Alr1/2
100 10 1
MurI D-Glu
L-Lys
AC C
EP
TE D
L-Glu
M AN U
0.1
SC
1000000
RI PT
ACCEPTED MANUSCRIPT
ACCEPTED MANUSCRIPT Cytoplasmic Peptidoglycan Intermediate Levels in S. aureus
Cytoplasmic Peptidoglycan Intermediate Levels in Staphylococcus aureus* Harika Vemula, Navid J. Ayon, and William G. Gutheil
RI PT
From the Division of Pharmaceutical Sciences, School of Pharmacy, University of Missouri-Kansas City, 2464 Charlotte Street, Kansas City, MO 64108 *Running title: Cytoplasmic Peptidoglycan Intermediate Levels in S. aureus
To whom correspondence should be addressed: William G. Gutheil, Division of Pharmaceutical Sciences, School of Pharmacy, University of Missouri Kansas City, 2464 Charlotte Street., Kansas City, MO, 64108, USA, Tel.: (816) 235-2424; Fax: (816) 235-5779; E- mail: [email protected]
M AN U
SC
Keywords: Peptidoglycan; Staphylococcus aureus; Metabolomics; Cell wall.
Highlights: • UDP-linked and amino acid peptidoglycan (PG) intermediates in S. aureus were quantified by LCMS/MS analysis. • An LC-MS/MS approach to total PG quantification was also developed, and used to estimate an S. aureus PG pathway flux of 690 µM/min. • S. aureus PG intermediate levels were compared with S. aureus pathway enzyme characteristics – and E. coli levels and enzyme characteristics. • This study provides the first quantification of both UDP-linked and amine peptidoglycan intermediate levels in S. aureus.
TE D
branch range from >80% saturation for MurA, Z, and C, to
S0300-9084(15)00364-8
DOI:
10.1016/j.biochi.2015.11.017
Reference:
BIOCHI 4884
To appear in:
Biochimie
Received Date: 11 May 2015 Accepted Date: 18 November 2015
Please cite this article as: H. Vemula, N.J. Ayon, W.G. Gutheil, Cytoplasmic Peptidoglycan Intermediate Levels in Staphylococcus aureus, Biochimie (2015), doi: 10.1016/j.biochi.2015.11.017. This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
L-Ala MurA/Z
MurB
MurC
MurD
D-Ala
Ddl ATP
MurE
100000
D-Ala-D-Ala
MurF
PEP NADPH ATP ATP ATP ATP UDP-NAG UDP-Tri UDP-Penta UDP-EP UDP-NAM UDP-Mono UDP-Di
10000 1000
PG
µM
Alr1/2
100 10 1
MurI D-Glu
L-Lys
AC C
EP
TE D
L-Glu
M AN U
0.1
SC
1000000
RI PT
ACCEPTED MANUSCRIPT
ACCEPTED MANUSCRIPT Cytoplasmic Peptidoglycan Intermediate Levels in S. aureus
Cytoplasmic Peptidoglycan Intermediate Levels in Staphylococcus aureus* Harika Vemula, Navid J. Ayon, and William G. Gutheil
RI PT
From the Division of Pharmaceutical Sciences, School of Pharmacy, University of Missouri-Kansas City, 2464 Charlotte Street, Kansas City, MO 64108 *Running title: Cytoplasmic Peptidoglycan Intermediate Levels in S. aureus
To whom correspondence should be addressed: William G. Gutheil, Division of Pharmaceutical Sciences, School of Pharmacy, University of Missouri Kansas City, 2464 Charlotte Street., Kansas City, MO, 64108, USA, Tel.: (816) 235-2424; Fax: (816) 235-5779; E- mail: [email protected]
M AN U
SC
Keywords: Peptidoglycan; Staphylococcus aureus; Metabolomics; Cell wall.
Highlights: • UDP-linked and amino acid peptidoglycan (PG) intermediates in S. aureus were quantified by LCMS/MS analysis. • An LC-MS/MS approach to total PG quantification was also developed, and used to estimate an S. aureus PG pathway flux of 690 µM/min. • S. aureus PG intermediate levels were compared with S. aureus pathway enzyme characteristics – and E. coli levels and enzyme characteristics. • This study provides the first quantification of both UDP-linked and amine peptidoglycan intermediate levels in S. aureus.
TE D
branch range from >80% saturation for MurA, Z, and C, to
