Identification of the Actin and Plasminogen Binding Regions of Group B Streptococcal Phosphoglycerate Kinase

Boone, Tyler J.; Tyrrell, Gregory J.

doi:10.1074/jbc.m112.361261

Cited by 31 publications

(23 citation statements)

References 61 publications

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“…1, fragments 14 and 16). These motifs consist of clustered, positively charged amino acids that have been shown to have a role in binding to glycosaminoglycans 69,72 , actin 123 , and plasminogen 123 . Heparin mimics the glycosaminoglycans found in the extracellular matrix and on the surface of host cells 124 .…”

Section: Discussionmentioning

confidence: 99%

“…Proline-rich regions in proteins have been implicated in protein:protein interactions [131][132][133] and it has been suggested that proline residues could anchor the C-terminus of P1 in the cell membrane 49 . Lysine-rich regions are associated with binding plasminogen 60,64,123,134,135 , heparin 59,61,69,72,115,136 , actin 116,123 , and DNA 75,137 . While P1-15 and P1-30 bound plasminogen in a dose-responsive manner, it was notable that RP-15 bound it more strongly.…”

Section: Discussionmentioning

confidence: 99%

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Cell surface processing of the P1 adhesin of Mycoplasma pneumoniae identifies novel domains that bind host molecules

Widjaja

Berry

Jarocki

et al. 2020

Sci Rep

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Mycoplasma pneumoniae is a genome reduced pathogen and causative agent of community acquired pneumonia. The major cellular adhesin, P1, localises to the tip of the attachment organelle forming a complex with P40 and P90, two cleavage fragments derived by processing Mpn142, and other molecules with adhesive and mobility functions. LC-MS/MS analysis of M. pneumoniae M129 proteins derived from whole cell lysates and eluents from affinity matrices coupled with chemically diverse host molecules identified 22 proteoforms of P1. Terminomics was used to characterise 17 cleavage events many of which were independently verified by the identification of semitryptic peptides in our proteome studies and by immunoblotting. One cleavage event released 1597 tSAAKpGApRppVppKpGApKppVQppKKpA 1627 from the C-terminus of P1 and this peptide was shown to bind to a range of host molecules. A smaller synthetic peptide comprising the C-terminal 15 amino acids, 1613 pGApKppVQppKKpA 1627 , selectively bound cytoskeletal intermediate filament proteins cytokeratin 7, cytokeratin 8, cytokeratin 18, and vimentin from a native A549 cell lysate. Collectively, our data suggests that ectodomain shedding occurs on the surface of M. pneumoniae where it may alter the functional diversity of P1, Mpn142 and other surface proteins such as elongation factor Tu via a mechanism similar to that described in Mycoplasma hyopneumoniae. The attachment organelle is a structurally and functionally sophisticated component of the M. pneumoniae cell that is responsible for the assembly of proteins essential for motility and adherence 1-8. An extensive list of host molecules including fibronectin 9-13 , fibrinogen 10-14 , plasminogen 11-13,15-17 , lactoferrin 10-12 , laminin 10-12 , and vitronectin 10-13 interact with surface accessible adhesins in M. pneumoniae. Other less well-defined host molecules include sialylated molecules 18 , oligosaccharides 19 , glycolipids 20 , and glycoproteins 21. The gene mpn141 encoding the major adhesin P1 is located in the same operon along with mpn140 and mpn142 and these three genes constitute a polycistronic transcriptional unit 22,23. mpn140 encodes for a 28 kDa putative phosphoesterase 24 and while it has been shown to degrade nanoRNA and dephosphorylate 3′-phosphoadenosine 5′-phosphate to AMP 25 , no role in adherence has been assigned for this protein. mpn142 generates a 130 kDa product (Mpn142) that is cleaved into two fragments of 40 kDa (P40) and 90 kDa (P90) immediately after or concurrent with translation 26,27. The cleavage event in Mpn142, first described over 25 years ago, was the first in what is now known to be a highly processed molecule on the surface of M. pneumoniae 28. P1 is a remarkably versatile molecule and the subject of numerous studies over the past 30 years. The only cleavage event that has been accurately assigned to P1 is the removal of the N-terminal 59 amino acids as a leader peptide 29. Molecular cross-linking and immunogold-labelling studies indicated that P1 forms a complex with P30, P40, and P90 ...

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Cell surface processing of the P1 adhesin of Mycoplasma pneumoniae identifies novel domains that bind host molecules

Widjaja

Berry

Jarocki

et al. 2020

Sci Rep

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“…Positively charged amino acids in SLiMs play a crucial role in interactions between proteins and highly sulphated glycosaminoglycans such as heparin 110 , and other molecules such as actin 111 , plasminogen 112 , DNA 113 , 114 and fibronectin 69 , 84 , 115 . Here we identified SLiMs enriched in positively charged amino acids in different regions of Mpn Ef-Tu , including sequences 37 aKegKsaatRy 47 , 183 pKweaKiHd 191 , and 248 lRpiRKa 254 , and identified eight surface exposed PPI sites, including three that reside within putative heparin-binding motifs 2 a Re KfdRsKpHv 13 , 73 d KRHyaHv 80 , and 370 e K gsKfsiReggRt 383 .…”

Section: Discussionmentioning

confidence: 99%

Elongation factor Tu is a multifunctional and processed moonlighting protein

Widjaja

Harvey

Hagemann

et al. 2017

Sci Rep

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Many bacterial moonlighting proteins were originally described in medically, agriculturally, and commercially important members of the low G + C Firmicutes. We show Elongation factor Tu (Ef-Tu) moonlights on the surface of the human pathogens Staphylococcus aureus (SaEf-Tu) and Mycoplasma pneumoniae (MpnEf-Tu), and the porcine pathogen Mycoplasma hyopneumoniae (MhpEf-Tu). Ef-Tu is also a target of multiple processing events on the cell surface and these were characterised using an N-terminomics pipeline. Recombinant MpnEf-Tu bound strongly to a diverse range of host molecules, and when bound to plasminogen, was able to convert plasminogen to plasmin in the presence of plasminogen activators. Fragments of Ef-Tu retain binding capabilities to host proteins. Bioinformatics and structural modelling studies indicate that the accumulation of positively charged amino acids in short linear motifs (SLiMs), and protein processing promote multifunctional behaviour. Codon bias engendered by an A + T rich genome may influence how positively-charged residues accumulate in SLiMs.

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“…The resulting surface-bound protease activity promotes disruption of host barriers thereby contributing to bacterial dissemination and invasiveness (Magalhaes et al, 2013;Six et al, 2015). Plg and Pln binding at the GBS surface is mediated by moonlighting proteins, such as a-enolase, glyceraldehyde dehydrogenase and phosphoglycerate kinase, which reassociate with the cell surface following bacterial lysis (Magalhaes et al, 2007;Boone and Tyrrell, 2012;Oliveira et al, 2012). Depending on their phylogenetic lineage, GBS strains also express unrelated fibrinogen receptors (FbsA, FbsB or FbsC) that catalyse indirect Plg binding (Gutekunst et al, 2004;Pietrocola et al, 2005;Buscetta et al, 2014).…”

Section: Introductionmentioning

confidence: 99%

PbsP, a cell wall‐anchored protein that binds plasminogen to promote hematogenous dissemination of group B Streptococcus

Buscetta

Firon

Pietrocola

et al. 2016

Molecular Microbiology

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Streptococcus agalactiae (Group B Streptococcus or GBS) is a leading cause of invasive infections in neonates whose virulence is dependent on its ability to interact with cells and host components. We here characterized a surface protein with a critical function in GBS pathophysiology. This adhesin, designated PbsP, possesses two Streptococcal Surface Repeat domains, a methionine and lysine-rich region, and a LPXTG cell wall-anchoring motif. PbsP mediates plasminogen (Plg) binding both in vitro and in vivo and we showed that cell surface-bound Plg can be activated into plasmin by tissue plasminogen activator to increase the bacterial extracellular proteolytic activity. Absence of PbsP results in a decreased bacterial transmigration across brain endothelial cells and impaired virulence in a murine model of infection. PbsP is conserved among the main GBS lineages and is a major plasminogen adhesin in non-CC17 GBS strains. Importantly, immunization of mice with recombinant PbsP confers protective immunity. Our results indicate that GBS have evolved different strategies to recruit Plg which indicates that the ability to acquire cell surface proteolytic activity is essential for the invasiveness of this bacterium.

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Identification of the Actin and Plasminogen Binding Regions of Group B Streptococcal Phosphoglycerate Kinase

Cited by 31 publications

References 61 publications

Cell surface processing of the P1 adhesin of Mycoplasma pneumoniae identifies novel domains that bind host molecules

Cell surface processing of the P1 adhesin of Mycoplasma pneumoniae identifies novel domains that bind host molecules

Elongation factor Tu is a multifunctional and processed moonlighting protein

PbsP, a cell wall‐anchored protein that binds plasminogen to promote hematogenous dissemination of group B Streptococcus

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