Changes in the protein profiles of planktonic cultures and biofilms of Staphylococcus epidermidis under anaerobic conditions in the presence of the hormone CNP

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Abstract

Natriuretic peptides (NP) are able to affect biofilms of human commensal microorganisms, including representatives of the genus Staphylococcus, however, the literature lacks data on the molecular changes caused by these hormones at the posttranslational level. In this regard, the present work shows for the first time that C-type natriuretic peptide (CNP) induces large changes in protein profiles of Staphylococcus aureus cells and biofilms. The presence of the hormone leads to a more pronounced difference in protein profiles between planktonic cells and biofilms when compared to control pairs of samples. The main processes affected are TCA cycle, protein transport, purine synthesis (decrease in the amount of the corresponding proteins in biofilms) and nitrate metabolism (increase in the amount of nitrogenases and other proteins in biofilms). It is necessary to mention separately the decreased amount of lysostaphin in biofilms compared to planktonic cultures when exposed to CNP. This may be one of the potential mechanisms of the recently shown reduction of competitive properties of S. epidermidis in the community with other microorganisms, which is induced by the presence of CNP in the medium. In addition, the results of the study strengthen the hypothesis that, as in the case of other human hormones, the action of CNP on S. epidermidis , is multitargeted. One of the likely mechanisms of the hormone’s action may be the disruption of the transition from planktonic culture to biofilm, which can be assumed without suppressing cell growth, which needs further verification.

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About the authors

A. V. Gannesen

FRC “Fundamentals of biotechnology” of RAS

Author for correspondence.
Email: andrei.gannesen@gmail.com
Russian Federation, Moscow, 119071

R. H. Ziganshin

Shemyakin-Ovchinnikov Institute of bioorganic chemistry of RAS

Email: andrei.gannesen@gmail.com
Russian Federation, Moscow, 117997

M. A. Ovcharova

FRC “Fundamentals of biotechnology” of RAS

Email: andrei.gannesen@gmail.com
Russian Federation, Moscow, 119071

A. M. Mosolova

FRC “Fundamentals of biotechnology” of RAS

Email: andrei.gannesen@gmail.com
Russian Federation, Moscow, 119071

N. A. Loginova

FRC “Fundamentals of biotechnology” of RAS

Email: andrei.gannesen@gmail.com
Russian Federation, Moscow, 119071

E. V. Diuvenji

FRC “Fundamentals of biotechnology” of RAS

Email: andrei.gannesen@gmail.com
Russian Federation, Moscow, 119071

E. D. Nevolina

FRC “Fundamentals of biotechnology” of RAS

Email: andrei.gannesen@gmail.com
Russian Federation, Moscow, 119071

S. V. Mart’yanov

FRC “Fundamentals of biotechnology” of RAS

Email: andrei.gannesen@gmail.com
Russian Federation, Moscow, 119071

V. K. Plakunov

FRC “Fundamentals of biotechnology” of RAS

Email: andrei.gannesen@gmail.com
Russian Federation, Moscow, 119071

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2. Fig. 1. Changes in the protein profiles of planktonic cultures depending on the cultivation time. A, B - control samples without the addition of the hormone CNP; C, D - samples with the addition of the hormone CNP. A, C - proteins in reduced quantities in 72-h planktonic cultures compared to 24-h cultures; B, D ‒ proteins in increased quantities in 72-h planktonic cultures compared to 24-h cultures. 1 - cluster of putative Zn-dependent dehydrogenases and PhnB-like proteins; 2 - cluster of protein synthesis: ribosomal protein S9 and translation initiation factor IF-3; 3 - cluster of D-alanyl transporter and ribosomal protein L7/L12; 4 - cluster of protein excretion and, possibly, quorum sensing; 5 - cluster of lipid (triglyceride) metabolism; 6 - cluster of folate biosynthesis proteins. 7 – cluster of cyclophilin-like proteins of the MRA-NP family.

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3. Fig. 2. Changes in the protein profiles of S. epidermidis biofilms depending on the cultivation time. A, B – control samples without the addition of the CNP hormone; C, D – samples with the addition of the CNP hormone. A, C – proteins in reduced quantities in 72-h biofilms compared to 24-h biofilms; B, D ‒ proteins in increased quantities in 72-h biofilms compared to 24-h. 1 – protein excretion cluster; 2 – 3,4-dihydroxy-2-butanone-4-phosphate synthase/GTP cyclohydrolase II and guanosine monophosphate reductase cluster; 3 – protein excretion cluster.

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4. Fig. 3. Changes in the protein composition of S. epidermidis biofilms compared to planktonic cultures after 24 h of cultivation. A, B – control samples; C, D – samples in the presence of CNP. A, B – proteins in reduced amounts in biofilms compared to planktonic cultures; B, D – proteins in increased amounts in biofilms compared to planktonic cultures. 1 – cluster of protein and ribosome synthesis; 2 – cluster of threonine and arginine synthesis; 3 – cluster of TCA cycle proteins; 4 – cluster of antioxidant protection proteins; 5 – cluster of threonine, serine and glycine metabolism; 6 – proteins of the cell wall and division; 7 – cluster of proteins of purine and secondary metabolite synthesis; 8 – cluster of TCA cycle proteins; 9 – cluster of nitrate metabolism proteins.

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5. Fig. 4. Changes in the protein composition of S. epidermidis biofilms compared to planktonic cultures after 72 h of cultivation. A, B – control samples; C, D – samples in the presence of CNP. A, B – proteins in reduced amounts in biofilms compared to planktonic cultures; B, D – proteins in increased amounts in biofilms compared to planktonic cultures. 1 – cluster of 2-oxoisovalerate dehydrogenase E3 and glutamate dehydrogenase; 2 – cluster of 3,4-dihydroxy-2-butanone-4-phosphate synthase/GTP cyclohydrolase II and guanosine monophosphate reductase; 3 – cluster of quorum-sensing proteins based on phenol-soluble modulins; 4 – purine synthesis through folate; 5 – cluster of thiamine synthesis proteins; 6 – cluster of glycerolipid metabolism. 7 – cluster of TCA cycle proteins; 8 – cluster of nitrate metabolism proteins.

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6. Fig. 5. Schematic diagram illustrating the effects of CNP on the protein composition of S. epidermidis planktonic cultures and biofilms, as well as the differences in protein profiles between planktonic cultures and staphylococcal biofilms.

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