Structural and functional insights into lysostaphin-substrate interaction

Helena Tossavainen, Vytas Raulinaitis, Linda Kauppinen, Ulla Pentikäinen, Hannu Maaheimo, Perttu Permi

Research output: Contribution to journalArticleScientificpeer-review

7 Citations (Scopus)

Abstract

Lysostaphin from Staphylococcus simulans and its family enzymes rapidly acquire prominence as the next generation agents in treatment of S. aureus infections. The specificity of lysostaphin is promoted by its C-terminal cell wall targeting domain selectivity toward pentaglycine bridges in S. aureus cell wall. Scission of these cross-links is carried out by its N-terminal catalytic domain, a zinc-dependent endopeptidase. Understanding the determinants affecting the efficiency of catalysis and strength and specificity of interactions lies at the heart of all lysostaphin family enzyme applications. To this end, we have used NMR, SAXS and molecular dynamics simulations to characterize lysostaphin structure and dynamics, to address the inter-domain interaction, the enzyme-substrate interaction as well as the catalytic properties of pentaglycine cleavage in solution. Our NMR structure confirms the recent crystal structure, yet, together with the molecular dynamics simulations, emphasizes the dynamic nature of the loops embracing the catalytic site. We found no evidence for inter-domain interaction, but, interestingly, the SAXS data delineate two preferred conformation subpopulations. Catalytic H329 and H360 were observed to bind a second zinc ion, which reduces lysostaphin pentaglycine cleaving activity. Binding of pentaglycine or its lysine derivatives to the targeting domain was found to be of very low affinity. The pentaglycine interaction site was located to the N-terminal groove of the domain. Notably, the targeting domain binds the peptidoglycan stem peptide Ala-D-γ-Glu-Lys-D-Ala-D-Ala with a much higher, micromolar affinity. Binding site mapping reveals two interaction sites of different affinities on the surface of the domain for this peptide.

Original languageEnglish
Article number60
JournalFrontiers in Molecular Biosciences
Volume5
Issue numberJUL
DOIs
Publication statusPublished - 3 Jul 2018
MoE publication typeNot Eligible

Fingerprint

Lysostaphin
Substrates
Molecular Dynamics Simulation
Cell Wall
Molecular dynamics
Zinc
Catalytic Domain
Enzymes
Cells
Nuclear magnetic resonance
Endopeptidases
Peptides
Peptidoglycan
Computer simulation
Catalysis
Staphylococcus
Lysine
Conformations
Crystal structure
Binding Sites

Keywords

  • Lysostaphin
  • NMR structure
  • Pentaglycine
  • Peptidoglycan
  • Protein dynamics
  • SH3b domain
  • Staphylococcus aureus
  • Substrate binding

Cite this

Tossavainen, Helena ; Raulinaitis, Vytas ; Kauppinen, Linda ; Pentikäinen, Ulla ; Maaheimo, Hannu ; Permi, Perttu. / Structural and functional insights into lysostaphin-substrate interaction. In: Frontiers in Molecular Biosciences. 2018 ; Vol. 5, No. JUL.
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Structural and functional insights into lysostaphin-substrate interaction. / Tossavainen, Helena; Raulinaitis, Vytas; Kauppinen, Linda; Pentikäinen, Ulla; Maaheimo, Hannu; Permi, Perttu.

In: Frontiers in Molecular Biosciences, Vol. 5, No. JUL, 60, 03.07.2018.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

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AU - Tossavainen, Helena

AU - Raulinaitis, Vytas

AU - Kauppinen, Linda

AU - Pentikäinen, Ulla

AU - Maaheimo, Hannu

AU - Permi, Perttu

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KW - Peptidoglycan

KW - Protein dynamics

KW - SH3b domain

KW - Staphylococcus aureus

KW - Substrate binding

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