scholarly journals The N2N3 domains of ClfA, FnbpA, and FnbpB in Staphylococcus aureus bind to human complement factor H, and their antibodies enhance the bactericidal capability of human blood

2020 ◽  
Author(s):  
Xinrui Mao ◽  
Junghyun Kim ◽  
QingFeng Zhang ◽  
TingTing Jiang ◽  
Dong Ho Ahn ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Cell Wall ◽  
Cell Surface ◽  
Human Blood ◽  
Bacterial Cell ◽  
Regulatory Protein ◽  
Factor H ◽  
Complement Factor ◽  
Bacterial Cell Surface ◽  
Protein Factor

Abstract In the complement system, the opsonin C3b binds to the bacterial cell surface and mediates the opsonophagocytosis. However, the cell wall protein SdrE of Staphylococcus aureus inhibits the C3b activity by recruiting the complement regulatory protein factor H (fH). SdrE binds to fH via its N-terminal N2N3 domain, which are also found in six other staphylococcal cell wall proteins. In this study, we report that not only the N2N3 domain of SdrE but also those of ClfA, FnbpA, and FnbpB can bind to fH. When immobilized on a microplate, the N2N3 domains recruited fH and enhanced the factor I (fI)-mediated cleavage of C3b. When mixed with fH and S. aureus cells, the N2N3 domains inhibited the fH binding to S. aureus cells and reduced the fI-mediated C3b cleavage on the bacterial cell surface. The F(ab)′2 fragments of the rabbit N2N3 antibodies also inhibited the fH-binding to the S. aureus cell surface. When added to human blood, the N2N3 antibodies or the N2N3 domain proteins significantly increased the bactericidal activity. Based on these results, we conclude that, in S. aureus, not only SdrE but also ClfA, FnbpA, and FnbpB can contribute to the inhibition of C3b-mediated opsonophagocytosis.

Sequence analysis of a cDNA clone encoding the C-terminal end of human complement factor H

1987 ◽  
Vol 7 (3) ◽  
pp. 201-207 ◽  
Author(s):  
A. J. Day ◽  
J. Ripoche ◽  
A. Lyons ◽  
B. McIntosh ◽  
T. J. R. Harris ◽  
...  
Keyword(s):  
Amino Acids ◽  
Tertiary Structure ◽  
Regulatory Protein ◽  
Peptide Sequencing ◽  
Factor H ◽  
Cdna Libraries ◽  
Complement Factor ◽  
Protein Factor ◽  
The Complement System ◽  
Full Length Clone

Peptide sequencing of the complement system regulatory protein, factor H, permitted the synthesis of a mixed sequence oligonucleotide probe. Human liver cDNA libraries were screened and factor H-specific clones selected. No full-length clone was obtained, but the largest available clone, R2a, was found to encode the C-terminal 657 amino acids of factor H. The derived amino acid sequence consists of 10 contiguous internally homologous segments, each about 60 amino acids long. Sequences homologous to these are found in several other complement and non-complement proteins. Such sequences are likely to represent a particular type of tertiary structure subunit.

scholarly journals Cleavage of Complement C3b to iC3b on the Surface of Staphylococcus aureus Is Mediated by Serum Complement Factor I

2004 ◽  
Vol 72 (5) ◽  
pp. 2858-2863 ◽  
Author(s):  
K. M. Cunnion ◽  
P. S. Hair ◽  
E. S. Buescher
Keyword(s):  
Staphylococcus Aureus ◽  
Human Serum ◽  
Serum Proteins ◽  
Regulatory Protein ◽  
Factor H ◽  
Complement Factor ◽  
Serum Factor ◽  
Complement Protein ◽  
Factor I ◽  
Serotype 5

ABSTRACT Complement-mediated opsonization of Staphylococcus aureus bearing the dominant capsule serotypes, serotypes 5 and 8, remains incompletely understood. We have previously shown that complement plays a vital role in the efficient phagocytosis of a serotype 5 S. aureus strain and that the opsonic fragments of the central complement protein C3, C3b and iC3b, were present on the bacterial surface after incubation in human serum. In the present studies, C3b and iC3b were found on several serotype 5 and 8 S. aureus strains after incubation in human serum. Using purified classical activation pathway complement proteins and the Western blot assay, we showed that when C3b was generated on the S. aureus surface no iC3b fragments were found, suggesting that other serum proteins may be required for cleaving C3b to iC3b. When C3b-coated S. aureus was incubated with serum factor I, a complement regulatory protein, iC3b was generated. Purified factor H, a serum protein cofactor for factor I, did not enhance factor I-mediated cleavage of C3b. These findings suggest that C3b cleavage to iC3b on S. aureus is mediated by serum factor I and does not require factor H.

scholarly journals M Protein of the Group A StreptococcusBinds to the Seventh Short Consensus Repeat of Human Complement Factor H

1998 ◽  
Vol 66 (4) ◽  
pp. 1427-1431 ◽  
Author(s):  
Timothy K. Blackmore ◽  
Vincent A. Fischetti ◽  
Tania A. Sadlon ◽  
Helena M. Ward ◽  
David L. Gordon
Keyword(s):  
Protein Binding ◽  
Binding Site ◽  
Regulatory Protein ◽  
Factor H ◽  
M Protein ◽  
Protein Binding Site ◽  
Enzyme Linked Immunosorbent Assay ◽  
Complement Factor ◽  
Heparin Binding ◽  
Protein Factor

ABSTRACT Streptococcus pyogenes evades complement by binding the complement-regulatory protein factor H (fH) via the central conserved C-repeat region of M protein. However, the corresponding binding region within fH has not previously been precisely localized. fH is composed of 20 conserved modules called short consensus repeats (SCRs), each of which contains approximately 60 amino acids. A series of fH truncated and deletion mutants were prepared, and their interaction with M6 protein was examined. The M protein binding site was initially localized to SCRs 6 to 15 as demonstrated by ligand dot blotting, chemical cross-linking, and enzyme-linked immunosorbent assay. SCR 7 was then shown to contain the M protein binding site, as a construct consisting of the first seven SCRs bound M protein but a construct containing the first six SCRs did not bind. In addition, deletion of SCR 7 from full-length fH abolished binding to M protein. SCR 7 is known to contain a heparin binding domain, and binding of fH to M6 protein was almost totally inhibited in the presence of 400 U of heparin per ml. These results localize the M6 protein binding site of fH to SCR 7 and indicate that it is in close proximity to the heparin binding site.

scholarly journals Clumping Factor A Interaction with Complement Factor I Increases C3b Cleavage on the Bacterial Surface of Staphylococcus aureus and Decreases Complement-Mediated Phagocytosis

2010 ◽  
Vol 78 (4) ◽  
pp. 1717-1727 ◽  
Author(s):  
Pamela S. Hair ◽  
Charlene G. Echague ◽  
Amber M. Sholl ◽  
Justin A. Watkins ◽  
Joan A. Geoghegan ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Surface Protein ◽  
Factor H ◽  
Complement Factor ◽  
Wild Type ◽  
Bacterial Surface ◽  
Complement Control Protein ◽  
Factor I ◽  
Protein Factor ◽  
Fibrinogen Binding

ABSTRACT The human complement system is important in the immunological control of Staphylococcus aureus infection. We showed previously that S. aureus surface protein clumping factor A (ClfA), when expressed in recombinant form, bound complement control protein factor I and increased factor I cleavage of C3b to iC3b. In the present study, we show that, compared to the results for the wild type, when isogenic ClfA-deficient S. aureus mutants were incubated in serum, they bound less factor I, generated less iC3b on the bacterial surface, and bound fewer C3 fragments. It has been shown previously that two amino acids in ClfA (P336 and Y338) are essential for fibrinogen binding. However, S. aureus expressing ClfA(P336A Y338S) was less virulent than ClfA-deficient strains in animal models. This suggested that ClfA contributed to S. aureus virulence by a mechanism different than fibrinogen binding. In the present study, we showed that S. aureus expressing ClfA(P336A Y338S) was more susceptible to complement-mediated phagocytosis than a ClfA-null mutant or the wild type. Unlike ClfA, ClfA(P336A Y338S) did not enhance factor I cleavage of C3b to iC3b and inhibited the cofactor function of factor H. Fibrinogen enhanced factor I binding to ClfA and the S. aureus surface. Twenty clinical S. aureus strains all expressed ClfA and bound factor I. High levels of factor I binding by clinical strains correlated with poor phagocytosis. In summary, our results suggest that the interaction of ClfA with factor I contributes to S. aureus virulence by a complement-mediated mechanism.

The structure of secondary cell wall polymers: how Gram-positive bacteria stick their cell walls together

Microbiology ◽  
2005 ◽  
Vol 151 (3) ◽  
pp. 643-651 ◽  
Author(s):  
Christina Schäffer ◽  
Paul Messner
Keyword(s):  
Cell Wall ◽  
Cell Surface ◽  
Bacterial Cell ◽  
Secondary Cell Wall ◽  
Chemical Study ◽  
Gram Positive ◽  
Gram Positive Bacteria ◽  
Bacterial Cell Surface ◽  
Cell Wall Polymers ◽  
Structural Principles

The cell wall of Gram-positive bacteria has been a subject of detailed chemical study over the past five decades. Outside the cytoplasmic membrane of these organisms the fundamental polymer is peptidoglycan (PG), which is responsible for the maintenance of cell shape and osmotic stability. In addition, typical essential cell wall polymers such as teichoic or teichuronic acids are linked to some of the peptidoglycan chains. In this review these compounds are considered as ‘classical’ cell wall polymers. In the course of recent investigations of bacterial cell surface layers (S-layers) a different class of ‘non-classical’ secondary cell wall polymers (SCWPs) has been identified, which is involved in anchoring of S-layers to the bacterial cell surface. Comparative analyses have shown considerable differences in chemical composition, overall structure and charge behaviour of these SCWPs. This review discusses the progress that has been made in understanding the structural principles of SCWPs, which may have useful applications in S-layer-based ‘supramolecular construction kits' in nanobiotechnology.

scholarly journals Tryptic Shaving of Staphylococcus aureus Unveils Immunodominant Epitopes on the Bacterial Cell Surface

2020 ◽  
Vol 19 (8) ◽  
pp. 2997-3010
Author(s):  
Annette Dreisbach ◽  
Min Wang ◽  
Magdalena M. van der Kooi-Pol ◽  
Ewoud Reilman ◽  
Dennis G. A. M. Koedijk ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Cell Surface ◽  
Bacterial Cell ◽  
Bacterial Cell Surface ◽  
Immunodominant Epitopes

scholarly journals A Quorum Sensing-Regulated Protein Binds Cell Wall Components and Enhances Lysozyme Resistance inStreptococcus pyogenes

2018 ◽  
Vol 200 (11) ◽  
Author(s):  
Artemis Gogos ◽  
Juan Cristobal Jimenez ◽  
Jennifer C. Chang ◽  
Reid V. Wilkening ◽  
Michael J. Federle
Keyword(s):  
Cell Wall ◽  
Quorum Sensing ◽  
Cell Surface ◽  
Bacterial Cell ◽  
Host Factor ◽  
Molecular Architecture ◽  
Gas Cell ◽  
Content Type ◽  
Bacterial Cell Surface ◽  
Positively Charged

ABSTRACTThe Rgg2/3 quorum sensing (QS) system is conserved among all sequenced isolates of group AStreptococcus(GAS;Streptococcus pyogenes). The molecular architecture of the system consists of a transcriptional activator (Rgg2) and a transcriptional repressor (Rgg3) under the control of autoinducing peptide pheromones (SHP2 and SHP3). Activation of the Rgg2/3 pathway leads to increases in biofilm formation and resistance to the bactericidal effects of the host factor lysozyme. In this work, we show that deletion of a small gene,spy49_0414c, abolished both phenotypes in response to pheromone signaling. The gene encodes a small, positively charged, secreted protein, referred to as StcA. Analysis of recombinant StcA showed that it can directly interact with GAS cell wall preparations containing phosphodiester-linked carbohydrate polymers but not with preparations devoid of them. Immunofluorescence microscopy detected antibody against StcA bound to the surface of paraformaldehyde-fixed wild-type cells. Expression of StcA in bacterial culture induced a shift in the electrostatic potential of the bacterial cell surface, which became more positively charged. These results suggest that StcA promotes phenotypes by way of ionic interactions with the GAS cell wall, most likely with negatively charged cell wall-associated polysaccharides.IMPORTANCEThis study focuses on a small protein, StcA, that is expressed and secreted under induction of Rgg2/3 QS, ionically associating with negatively charged domains on the cell surface. These data present a novel mechanism of resistance to the host factor lysozyme by GAS and have implications in the relevance of this circuit in the interaction between the bacterium and the human host that is mediated by the bacterial cell surface.

scholarly journals Lectin Microarray Reveals Binding Profiles of Lactobacillus casei Strains in a Comprehensive Analysis of Bacterial Cell Wall Polysaccharides

2011 ◽  
Vol 77 (13) ◽  
pp. 4539-4546 ◽  
Author(s):  
Emi Yasuda ◽  
Hiroaki Tateno ◽  
Jun Hirabarashi ◽  
Tohru Iino ◽  
Tomoyuki Sako
Keyword(s):  
Cell Wall ◽  
Cell Surface ◽  
Bacterial Cell ◽  
Lactobacillus Casei ◽  
Lectin Binding ◽  
Bacterial Strains ◽  
Cell Wall Polysaccharides ◽  
Content Type ◽  
Lectin Microarray ◽  
Bacterial Cell Surface

ABSTRACTWe previously showed a pivotal role of the polysaccharide (PS) moiety in the cell wall of theLactobacillus caseistrain Shirota (YIT 9029) as a possible immune modulator (E. Yasuda M. Serata, and T. Sako, Appl. Environ. Microbiol. 74:4746-4755, 2008). To distinguish PS structures on the bacterial cell surface of individual strains in relation to their activities, it would be useful to have a rapid and high-throughput methodology. Recently, a new technique called lectin microarray was developed for rapid profiling of glycosylation in eukaryotic polymers and cell surfaces. Here, we report on the development of a simple and sensitive method based on this technology for direct analysis of intact bacterial cell surface glycomes. The method involves labeling bacterial cells with SYTOX Orange before incubation with the lectin microarray. After washing, bound cells are directly detected using an evanescent-field fluorescence scanner in a liquid phase. Using this method, we compared the cell surface glycomes from 16 different strains ofL. casei. The patterns of lectin-binding affinity of most strains were found to be unique. There appears to be two types of lectin-binding profiles: the first is characterized by a few lectins, and the other is characterized by multiple lectins with different specificities. We also showed a dramatic change in the lectin-binding profile of a YIT 9029 derivative with a mutation in thecps1Cgene, encoding a putative glycosyltransferase. In conclusion, the developed technique provided a novel strategy for rapid profiling and, more importantly, differentiating numerous bacterial strains with relevance to the biological functions of PS.

scholarly journals Human complement factor H deficiency associated with hemolytic uremic syndrome.

1998 ◽  
Vol 9 (12) ◽  
pp. 2318-2326
Author(s):  
N Rougier ◽  
M D Kazatchkine ◽  
J P Rougier ◽  
V Fremeaux-Bacchi ◽  
J Blouin ◽  
...  
Keyword(s):  
Hemolytic Uremic Syndrome ◽  
Regulatory Protein ◽  
Large Deletion ◽  
Factor H ◽  
Pcr Analysis ◽  
Complement Factor ◽  
Human Complement ◽  
Protein Factor ◽  
Factor H Deficiency ◽  
Uremic Syndrome

This study reports on six cases of deficiency in the human complement regulatory protein Factor H (FH) in the context of an acute renal disease. Five of the cases were observed in children presenting with idiopathic hemolytic uremic syndrome (HUS). Two of the children exhibited a homozygous deficiency characterized by the absence of the 150-kD form of Factor H and the presence, upon immunoblotting, of the 42-kD Factor H-like protein 1 (FHL-1) and other FH-related protein (FHR) bands. Southern blot and PCR analysis of DNA of one patient with homozygous deficiency ruled out the presence of a large deletion of the FH gene as the underlying defect for the deficiency. The other four children presented with heterozygous deficiency and exhibited a normal immunoblotting pattern of proteins of the FH family. Factor H deficiency is the only complement deficiency associated with HUS. These observations suggest a role for FH and/or FH receptors in the pathogenesis of idiopathic HUS.

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