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Protein & Peptide Letters

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ISSN (Print): 0929-8665
ISSN (Online): 1875-5305

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Research Article

Biophysical Characterization of Type III Pantothenate Kinase (PanK) from Acinetobacter baumannii

Author(s): Ankita Singla, Pradeep Sharma, Akshita Gupta, Naseer Iqbal , Chitra Rani, T.P. Singh and Sujata Sharma*

Volume 28, Issue 4, 2021

Published on: 13 August, 2020

Page: [450 - 458] Pages: 9

DOI: 10.2174/0929866527666200813202445

Price: $65

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Abstract

Background: Type-III Pantothenate kinase from the multi drug resistant bacteria, Acinetobacter baumannii (AbPanK) catalyzes the first step of the essential Coenzyme A biosynthesis pathway. AbPanK is an attractive drug target against the bacteria since it is an essential enzyme and its structure is significantly different from the human PanK.

Methods: AbPanK was cloned, expressed, purified and crystallized. A good quality single crystal was used for X-ray intensity data collection. Dynamic light scattering was done for calculating the hydrodynamic radii and its oligomeric nature in the solution. Binding studies of this protein with its two substrates, Pantothenate and ATP were done using spectrofluorometer.

Results: Our results indicated that AbPanK shows a strong affinity with pantothenate with dissociation constant of 1.2 x 10- 8 M and moderate affinity towards ATP of 3.7x 10-3 M. This fact was further substantiated by the calculations of Km of both substrates using kinase assay kit. Dynamic light scattering studies have shown that it exists as homogenous solution with hydrodynamic radii corresponding to the molecular weight of 29.55 kDa. A low-resolution X-ray intensity data set was collected, which shows that AbPank crystallizes in P2 space group with cell dimensions of a= 165 Å, b= 260 Å, and, c= 197 Å and α= 90.0, β= 113.60, γ= 90.0.

Discussion: Recombinant Pantothenate kinase from Acinetobacter baumannii was purified to homogeneity and crystallized. The enzyme exhibits very low sequence identity (28%) to other corresponding enzymes.

Conclusion: The recombinant enzyme was active and its binding affinities with its substrates pantothenate and ATP have been studied. This information would be very useful while designing the inhibitors of this enzyme in order to fight bacterial infections associated to this pathogen.

Keywords: Acinetobacter baumannii, pathogen, multidrug resistance, coaX, coenzyme A, pantothenate kinase.

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[1]
Maragakis, L.L.; Perl, T.M.; Perl, T.M. Acinetobacter baumannii: epidemiology, antimicrobial resistance, and treatment options. Clin. Infect. Dis., 2008, 46(8), 1254-1263.
[http://dx.doi.org/10.1086/529198] [PMID: 18444865]
[2]
Peleg, A.Y.; Seifert, H.; Paterson, D.L. Acinetobacter baumannii: emergence of a successful pathogen. Clin. Microbiol. Rev., 2008, 21(3), 538-582.
[http://dx.doi.org/10.1128/CMR.00058-07] [PMID: 18625687]
[3]
Santajit, S.; Indrawattana, N. Mechanisms of Antimicrobial Resistance in ESKAPE Pathogens. BioMed Res. Int., 2016, 20162475067
[http://dx.doi.org/10.1155/2016/2475067] [PMID: 27274985]
[4]
Abbo, A.; Carmeli, Y.; Navon-Venezia, S.; Siegman-Igra, Y.; Schwaber, M.J. Impact of multi-drug-resistant Acinetobacter baumannii on clinical outcomes. Eur. J. Clin. Microbiol. Infect. Dis., 2007, 26(11), 793-800.
[http://dx.doi.org/10.1007/s10096-007-0371-8] [PMID: 17701063]
[5]
Abbo, A.; Navon-Venezia, S.; Hammer-Muntz, O.; Krichali, T.; Siegman-Igra, Y.; Carmeli, Y. Multidrug-resistant Acinetobacter baumannii. Emerg. Infect. Dis., 2005, 11(1), 22-29.
[http://dx.doi.org/10.3201/eid1101.040001] [PMID: 15705318]
[6]
Rice, L.B. Federal funding for the study of antimicrobial resistance in nosocomial pathogens: no ESKAPE. J. Infect. Dis., 2008, 197(8), 1079-1081.
[http://dx.doi.org/10.1086/533452] [PMID: 18419525]
[7]
Scott, P.; Deye, G.; Srinivasan, A.; Murray, C.; Moran, K.; Hulten, E.; Fishbain, J.; Craft, D.; Riddell, S.; Lindler, L.; Mancuso, J.; Milstrey, E.; Bautista, C.T.; Patel, J.; Ewell, A.; Hamilton, T.; Gaddy, C.; Tenney, M.; Christopher, G.; Petersen, K.; Endy, T.; Petruccelli, B. An outbreak of multidrug-resistant Acinetobacter baumannii-calcoaceticus complex infection in the US military health care system associated with military operations in Iraq. Clin. Infect. Dis., 2007, 44(12), 1577-1584.
[http://dx.doi.org/10.1086/518170] [PMID: 17516401]
[8]
Kalanuria, A.A.; Ziai, W.; Mirski, M. Ventilator-associated pneumonia in the ICU. Crit. Care, 2014, 18(2), 208.
[http://dx.doi.org/10.1186/cc13775] [PMID: 25029020]
[9]
Manchanda, V.; Sanchaita, S.; Singh, N. Multidrug resistant acinetobacter. J. Glob. Infect. Dis., 2010, 2(3), 291-304.
[http://dx.doi.org/10.4103/0974-777X.68538] [PMID: 20927292]
[10]
Lee, C.R.; Lee, J.H.; Park, M.; Park, K.S.; Bae, I.K.; Kim, Y.B.; Cha, C.J.; Jeong, B.C.; Lee, S.H. Biology of Acinetobacter baumannii: Pathogenesis, Antibiotic Resistance Mechanisms, and Prospective Treatment Options. Front. Cell. Infect. Microbiol., 2017, 7, 55.
[http://dx.doi.org/10.3389/fcimb.2017.00055] [PMID: 28348979]
[11]
Bassetti, M.; Righi, E.; Esposito, S.; Petrosillo, N.; Nicolini, L. Drug treatment for multidrug-resistant Acinetobacter baumannii infections. Future Microbiol., 2008, 3(6), 649-660.
[http://dx.doi.org/10.2217/17460913.3.6.649] [PMID: 19072182]
[12]
Spry, C.; Kirk, K.; Saliba, K.J. Coenzyme A biosynthesis: an antimicrobial drug target. FEMS Microbiol. Rev., 2008, 32(1), 56-106.
[http://dx.doi.org/10.1111/j.1574-6976.2007.00093.x] [PMID: 18173393]
[13]
Song, W.J.; Jackowski, S. Cloning, sequencing, and expression of the pantothenate kinase (coaA) gene of Escherichia coli. J. Bacteriol., 1992, 174(20), 6411-6417.
[http://dx.doi.org/10.1128/JB.174.20.6411-6417.1992] [PMID: 1328157]
[14]
Nicely, N.I.; Parsonage, D.; Paige, C.; Newton, G.L.; Fahey, R.C.; Leonardi, R.; Jackowski, S.; Mallett, T.C.; Claiborne, A. Structure of the type III pantothenate kinase from Bacillus anthracis at 2.0 A resolution: implications for coenzyme A-dependent redox biology. Biochemistry, 2007, 46(11), 3234-3245.
[http://dx.doi.org/10.1021/bi062299p] [PMID: 17323930]
[15]
Leonardi, R.; Jackowski, S. Biosynthesis of Pantothenic Acid and Coenzyme A. Ecosal Plus, 2007, 2(2), 1128.
[http://dx.doi.org/10.1128/ecosalplus.3.6.3.4] [PMID: 26443589]
[16]
Brand, L.A.; Strauss, E. Characterization of a new pantothenate kinase isoform from Helicobacter pylori. J. Biol. Chem., 2005, 280(21), 20185-20188.
[http://dx.doi.org/10.1074/jbc.C500044200] [PMID: 15795230]
[17]
Leonardi, R.; Chohnan, S.; Zhang, Y.M.; Virga, K.G.; Lee, R.E.; Rock, C.O.; Jackowski, S. A pantothenate kinase from Staphylococcus aureus refractory to feedback regulation by coenzyme A. J. Biol. Chem., 2005, 280(5), 3314-3322.
[http://dx.doi.org/10.1074/jbc.M411608200] [PMID: 15548531]
[18]
Song, W.J.; Jackowski, S. Kinetics and regulation of pantothenate kinase from Escherichia coli. J. Biol. Chem., 1994, 269(43), 27051-27058.
[http://dx.doi.org/10.1016/S0021-9258(18)47124-4] [PMID: 7929447]
[19]
Yang, K.; Strauss, E.; Huerta, C.; Zhang, H. Structural basis for substrate binding and the catalytic mechanism of type III pantothenate kinase. Biochemistry, 2008, 47(5), 1369-1380.
[http://dx.doi.org/10.1021/bi7018578] [PMID: 18186650]
[20]
Hong, B.S.; Yun, M.K.; Zhang, Y.M.; Chohnan, S.; Rock, C.O.; White, S.W.; Jackowski, S.; Park, H.W.; Leonardi, R. Prokaryotic type II and type III pantothenate kinases: The same monomer fold creates dimers with distinct catalytic properties. Structure, 2006, 14(8), 1251-1261.
[http://dx.doi.org/10.1016/j.str.2006.06.008] [PMID: 16905099]
[21]
Battye, T.G.G.; Kontogiannis, L.; Johnson, O.; Powell, H.R.; Leslie, A.G.W. iMOSFLM: a new graphical interface for diffraction-image processing with MOSFLM. Acta Crystallogr. D Biol. Crystallogr., 2011, 67(Pt 4), 271-281.
[http://dx.doi.org/10.1107/S0907444910048675] [PMID: 21460445]
[22]
Yang, K.; Eyobo, Y.; Brand, L.A.; Martynowski, D.; Tomchick, D.; Strauss, E.; Zhang, H. Crystal structure of a type III pantothenate kinase: insight into the mechanism of an essential coenzyme A biosynthetic enzyme universally distributed in bacteria. J. Bacteriol., 2006, 188(15), 5532-5540.
[http://dx.doi.org/10.1128/JB.00469-06] [PMID: 16855243]
[23]
Vincent, J.L.; Rello, J.; Marshall, J.; Silva, E.; Anzueto, A.; Martin, C.D.; Moreno, R.; Lipman, J.; Gomersall, C.; Sakr, Y.; Reinhart, K. International study of the prevalence and outcomes of infection in intensive care units. JAMA, 2009, 302(21), 2323-2329.
[http://dx.doi.org/10.1001/jama.2009.1754] [PMID: 19952319]
[24]
Boucher, H.W.; Talbot, G.H.; Bradley, J.S.; Edwards, J.E.; Gilbert, D.; Rice, L.B.; Scheld, M.; Spellberg, B.; Bartlett, J. Bad bugs, no drugs: no ESKAPE! An update from the Infectious Diseases Society of America. Clin. Infect. Dis., 2009, 48(1), 1-12.
[http://dx.doi.org/10.1086/595011] [PMID: 19035777]

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