Biochemical Characterization of Recombinant Fusions of Lipopolysaccharide Binding Protein and Bactericidal/Permeability-increasing Protein

Abrahamson, Susan L.; Wu, Hung Tsung; Williams, Robert E.; Der, Ken; Ottah, Nneka; Little, R.; Gazzano-Santoro, Hélène; Theofan, Georgia; Bauer, Robert J.; Leigh, Scott; Orme, Anne; Horwitz, Arnold H.; Carroll, Stephen F.; Dedrick, Russell L.

doi:10.1074/jbc.272.4.2149

Cited by 74 publications

(82 citation statements)

References 40 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In contrast to the effect of binding of LBP to endotoxin, binding of BPI does not lead to extraction and transfer of endotoxin to CD14 and thus can preclude MD-2/TLR4-dependent cell activation by endotoxin [51,66]. This difference reflects differences in the structural and functional properties of the C-terminal domain of LBP and of BPI [48,51,81,107]. Additional differences in the structural and functional properties of the N-terminal domain of BPI and of LBP account for the higher affinity of BPI (vs. LBP) for purified or outer membrane-inserted endotoxin [9,52,53].…”

Section: Bpi Actions During Host: Gnb Interactionsmentioning

confidence: 96%

“…Basic residues clustered at the far end of the amino-terminal domain of BPI are thought to mediate initial electrostatic interactions with acidic sites of endotoxin located close to the lipid A region [47]. In addition, apolar pockets were demonstrated on the concave side of the molecule where host-derived phospholipids can be bound [46] that may be needed to stabilize the water soluble folded structure of BPI [46,48]. Studies using the recombinant amino-terminal domain of BPI alone or recombinant human BPI N /LBP C and LBP N /BPI C chimeric fusion proteins [48,49] have confirmed that the N-terminal domain (minimal structure of residues 13-191; [50]) is responsible for the antimicrobial cytotoxicity [45] and endotoxin-neutralizating properties of BPI [9,49] whereas the C-terminal domain of BPI is needed for BPI-dependent delivery of intact GNB and cell-fee endotoxin-rich particles to specific host cells [51][52][53].…”

Section: Bpi: Structure and Functionmentioning

confidence: 99%

“…In addition, apolar pockets were demonstrated on the concave side of the molecule where host-derived phospholipids can be bound [46] that may be needed to stabilize the water soluble folded structure of BPI [46,48]. Studies using the recombinant amino-terminal domain of BPI alone or recombinant human BPI N /LBP C and LBP N /BPI C chimeric fusion proteins [48,49] have confirmed that the N-terminal domain (minimal structure of residues 13-191; [50]) is responsible for the antimicrobial cytotoxicity [45] and endotoxin-neutralizating properties of BPI [9,49] whereas the C-terminal domain of BPI is needed for BPI-dependent delivery of intact GNB and cell-fee endotoxin-rich particles to specific host cells [51][52][53]. Reported anti-angiogenic properties have been described thus far for the BPI holoprotein and for a recombinant N-terminal fragment of BPI (residues 1-191; rBPI 21 [54][55][56]) whereas enhancement of complement activation on bacterial surfaces has been described only for the full-length BPI molecule [57].…”

Section: Bpi: Structure and Functionmentioning

confidence: 99%

“…Although BPI binding to aggregates of endotoxin, cell-free outer membrane blebs or intact GNB does not engage CD14-dependent cellular interactions, BPI can promote CD14-independent interactions with specific host cells [48,[51][52][53]. In each instance, these effects of BPI are dependent on the N-teminal domain of BPI to bind endotoxin and the C-terminal domain of BPI to somehow facilitate targeting of the BPI-coated endotoxin-rich particle to a host cell.…”

Section: Bpi Actions During Host: Gnb Interactionsmentioning

confidence: 99%

See 3 more Smart Citations

The bactericidal/permeability-increasing protein (BPI) in infection and inflammatory disease

Schultz

Weiss

2007

Clinica Chimica Acta

114

View full text Add to dashboard Cite

Gram-negative bacteria (GNB) and their endotoxin present a constant environmental challenge. Endotoxins can potently signal mobilization of host defenses against invading GNB but also potentially induce severe pathophysiology, necessitating controlled initiation and resolution of endotoxin-induced inflammation to maintain host integrity. The bactericidal/permeability-increasing protein (BPI) is a pluripotent protein expressed, in humans, mainly neutrophils. BPI exhibits strong anti-microbial activity against GNB and potent endotoxin-neutralizing activity. BPI mobilized with neutrophils in response to invading GNB can promote intracellular and extracellular bacterial killing, endotoxin neutralization and clearance, and delivery of GNB outer membrane antigens to dendritic cells. Tissue expression by dermal fibroblasts and epithelia could further amplify local levels of BPI and local interaction with GNB and endotoxin, helping to constrain local tissue infection and inflammation and prevent systemic infection and systemic inflammation. This review article focuses on the structural and functional properties of BPI with respect to its contribution to host defense during GNB infections and endotoxin-induced inflammation and the genesis of auto-antibodies against BPI that can blunt BPI activity and potentially contribute to chronic inflammatory disease.

show abstract

Section: Bpi Actions During Host: Gnb Interactionsmentioning

confidence: 96%

Section: Bpi: Structure and Functionmentioning

confidence: 99%

Section: Bpi: Structure and Functionmentioning

confidence: 99%

Section: Bpi Actions During Host: Gnb Interactionsmentioning

confidence: 99%

See 2 more Smart Citations

The bactericidal/permeability-increasing protein (BPI) in infection and inflammatory disease

Schultz

Weiss

2007

Clinica Chimica Acta

114

View full text Add to dashboard Cite

show abstract

“…In this study, LBP, BPI, and previously described chimeric proteins of LBP and BPI (39) have been utilized with metabolically labeled preparations of Escherichia coli K12 LPS (40) to further define the role of the COOH-terminal domain of BPI and LBP in delivery of endotoxin to specific cellular and extracellular host targets. We compared the effects of these proteins on the binding of LPS to and activation of enriched populations of leukocytes and on the interaction of LPS with sCD14.…”

mentioning

confidence: 99%

The Carboxyl-terminal Domain of Closely Related Endotoxin-binding Proteins Determines the Target of Protein-Lipopolysaccharide Complexes

Iovine¹,

Eastvold²,

Elsbach³

et al. 2002

Journal of Biological Chemistry

View full text Add to dashboard Cite

Bacterial invasion triggers sensitive, specific molecular alarms that result in the mobilization of host defenses designed to recognize and eliminate invading bacteria and their remnants. The major inducer of host responses to Gram-negative bacteria (GNB) 1 is endotoxin, i.e. lipopolysaccharide, LPS, the unique glycolipid that comprises the bulk of the outer leaflet of the GNB outer membrane (1). These amphipathic molecules are composed of a unique, conserved hydrophobic moiety, the lipid A region, that is a disaccharide of N-acetylglucosamine substituted with saturated fatty acids and attached to a highly charged acidic carbohydrate region of varying size and composition (1, 2). The chemical structure of endotoxin promotes the formation of highly ordered, but potentially malleable aggregated state(s). Interaction with host endotoxin-binding proteins alters the physical presentation of endotoxin and its ability to activate cell responses (2-5). The cumulative work of many laboratories has implicated lipopolysaccharide-binding protein (LBP), CD14, Toll-like receptor 4, and MD-2 as key factors in cell activation by LPS (3, 6 -13). LBP facilitates delivery of LPS to both membrane-bound, GPI-linked (mCD14) and soluble CD14 (sCD14) (14 -19). Through interaction with CD14, LPS activates cells via a transmembrane receptor capable of promoting signal transduction. This recognition/response cascade includes the Toll-like receptor family of proteins, most notably Toll-like receptor 4, that serve as the primary mediator of endotoxin signaling (20, 21).The host also utilizes defense mechanisms that blunt endotoxin-triggered inflammatory responses by eliminating viable GNB and by neutralizing endotoxin. One potent host protein that blocks LPS activity is bactericidal/permeability increasing protein (BPI), a basic protein residing in azurophilic granules of polymorphonuclear (PMN) leukocytes and in the extracellular fluid of PMN-rich inflammatory exudates (22). BPI efficiently neutralizes LPS and is also potently cytotoxic and opsonic, especially toward GNB (22)(23)(24)(25). Although the functional properties of LBP and BPI differ markedly, they share 45% amino acid identity, are encoded within the same region of chromosome 20, and belong to a family of lipid-binding proteins that include phospholipid transfer protein and cholesteryl ester transfer protein (26 -30). The three-dimensional crystal structure for BPI reveals an unusual "hinged" two-domain boomerang-like molecular structure. The extensive sequence homology between LBP and BPI predicts a nearly superimposable threedimensional structure for LBP (30) suggesting that despite their different bioactivities, BPI and LBP have a similar organization of structure and function.In support of this view, studies on the interaction of LBP and BPI with LPS have demonstrated that the NH 2 -terminal domain of each protein is responsible for binding LPS (28,(31)(32)(33)(34). In BPI, both affinity for LPS and antibacterial activity are concentrated in this portion of the protein (31-...

show abstract

Design, Synthesis and Characterization of Cationic Peptide and Steroid Antibiotics

Savage

2002

Eur. J. Org. Chem.

View full text Add to dashboard Cite

Biochemical Characterization of Recombinant Fusions of Lipopolysaccharide Binding Protein and Bactericidal/Permeability-increasing Protein

Cited by 74 publications

References 40 publications

The bactericidal/permeability-increasing protein (BPI) in infection and inflammatory disease

The bactericidal/permeability-increasing protein (BPI) in infection and inflammatory disease

The Carboxyl-terminal Domain of Closely Related Endotoxin-binding Proteins Determines the Target of Protein-Lipopolysaccharide Complexes

Design, Synthesis and Characterization of Cationic Peptide and Steroid Antibiotics

Contact Info

Product

Resources

About