Structure-Function Relationships of the Complement Regulatory Protein, CD59

Petranka, John G.; Zhao, Jinmin; Norris, John D.; Tweedy, Neil; Ware, Russell E.; Sims, P.; Rosse, Wendell F.

doi:10.1006/bcmd.1996.0111

Cited by 44 publications

(62 citation statements)

References 43 publications

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“…Human CD59 contains 10 cysteines involved in 5 disulfide bridges that are critical for maintaining its three-dimensional conformation (35). Mutations that affect disulfide bridge formation in hCD59 result in low expression, loss of function, misfolding of CD59, and loss of all antibody epitopes (22,23,26,35).…”

Section: Resultsmentioning

confidence: 99%

Mapping the Intermedilysin-Human CD59 Receptor Interface Reveals a Deep Correspondence with the Binding Site on CD59 for Complement Binding Proteins C8α and C9

Wickham

Hotze

Farrand

et al. 2011

Journal of Biological Chemistry

106

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CD59 is a glycosylphosphatidylinositol-anchored protein that inhibits the assembly of the terminal complement membrane attack complex (MAC) pore, whereas Streptococcus intermedius intermedilysin (ILY), a pore forming cholesterol-dependent cytolysin (CDC), specifically binds to human CD59 (hCD59) to initiate the formation of its pore. The identification of the residues of ILY and hCD59 that form their binding interface revealed a remarkably deep correspondence between the hCD59 binding site for ILY and that for the MAC proteins C8␣ and C9. ILY disengages from hCD59 during the prepore to pore transition, suggesting that loss of this interaction is necessary to accommodate specific structural changes associated with this transition. Consistent with this scenario, mutants of hCD59 or ILY that increased the affinity of this interaction decreased the cytolytic activity by slowing the transition of the prepore to pore but not the assembly of the prepore oligomer. A signature motif was also identified in the hCD59 binding CDCs that revealed a new hCD59-binding member of the CDC family. Although the binding site on hCD59 for ILY, C8␣, and C9 exhibits significant homology, no similarity exists in their binding sites for hCD59. Hence, ILY and the MAC proteins interact with common amino acids of hCD59 but lack detectable conservation in their binding sites for hCD59.The cholesterol-dependent cytolysins (CDCs) 4 are a family of pore-forming toxins produced by a diverse group of Grampositive pathogens. Recently crystal structures of the membrane attack complex/perforin proteins complement C8␣, a C9-like protein from Photorhabdus luminescens, and mouse perforin (1-4) suggested that they are structurally and mechanistically related the CDCs (5-7) and may be ancient ancestors (8). Interestingly, this relationship between the CDCs and membrane attack complex/perforin proteins extends to other features of the complement system. Two members of the CDC family, intermedilysin (ILY) secreted by Streptococcus intermedius and vaginolysin (VLY) secreted by Gardnerella vaginalis, specifically bind to the human form of CD59 (hCD59), a glycosylphosphatidylinositol (GPI)-anchored terminal inhibitor of the mammalian complement membrane attack complex (MAC) (9), rather than cholesterol (10, 11). These CDCs bind to hCD59 to initiate the assembly of their oligomeric pore complex on the membrane of human cells (10), whereas the main function of CD59 is to block the assembly of the MAC pore on host cells, thereby protecting them from the lytic effects of activated complement MAC. Domain 4 of the CDCs mediates its interaction with cholesterol-rich membranes (12)(13)(14). Most CDCs use cholesterol as their receptor and specifically recognize its 3-␤-hydroxy group. Recently Farrand et al. (15) defined the cholesterol recognition motif as a threonine/leucine pair located in loop 1 that is conserved in all members of the CDC family, including ILY and VLY. However, ILY initiates its interaction with the cell by binding to hCD59 (10) rather than chole...

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Section: Resultsmentioning

confidence: 99%

Mapping the Intermedilysin-Human CD59 Receptor Interface Reveals a Deep Correspondence with the Binding Site on CD59 for Complement Binding Proteins C8α and C9

Wickham

Hotze

Farrand

et al. 2011

Journal of Biological Chemistry

106

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“…Before embarking on our experiments, our fears appeared well founded; mutation of several of the cysteines within CD59 blocked trafficking of the molecule to the cell surface (13), and mutations of cysteines in the ligand-binding repeats of the LDL receptor have similar effects (18). However, as it turned out, mutations of cysteines in GPIHBP1 had only minor effects on cell surface expression of GPIHBP1.…”

Section: Discussionmentioning

confidence: 99%

“…For other Ly6 proteins, mutation of conserved cysteines appears to impair protein function. Mutation of several (but not all) of the Ly6 cysteines in CD59 reduces the ability of that molecule to inhibit complement-mediated membrane attack (13). Also, missense mutations involving cysteines were found in a secreted Ly6 protein, SLURP1, in association with a recessive palmoplantar keratoderma (16).…”

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confidence: 99%

“…But as we embarked on these studies, we worried that the cellbased assay system might be inadequate, for the simple reason that cysteine mutations sometimes interfere with the ability of proteins to reach the cell surface (13,18). Accordingly, we also developed a cell-free, monoclonal antibody-based assay system for assessing the ability of GPIHBP1 to bind LPL.…”

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confidence: 99%

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Highly Conserved Cysteines within the Ly6 Domain of GPIHBP1 Are Crucial for the Binding of Lipoprotein Lipase

Beigneux

Gin

Davies

et al. 2009

Journal of Biological Chemistry

138

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GPIHBP1, a glycosylphosphatidylinositol-anchored endothelial cell protein of the lymphocyte antigen 6 (Ly6) family, binds lipoprotein lipase (LPL) avidly and is required for the lipolytic processing of triglyceride-rich lipoproteins. GPIHBP1 contains two key structural motifs, an acidic domain and an Ly6 motif (a three-fingered domain specified by 10 cysteines). The acidic domain is required for LPL binding, but the importance of the Ly6 domain is less clear. To explore that issue, we transfected cells with a wild-type GPIHBP1 expression vector or mutant GPIHBP1 vectors in which specific cysteines in the Ly6 domain were changed to alanine. The mutant GPIHBP1 proteins reached the cell surface, as judged by antibody binding studies and by the ability of a phosphatidylinositol-specific phospholipase C to release these proteins from the cell surface. However, cells expressing the cysteine mutants could not bind LPL. The acidic domain of the cysteine mutants appeared to remain accessible, as judged by binding studies with an antibody against the acidic domain. We also developed a cell-free assay of LPL binding. We created a rat monoclonal antibody against the carboxyl terminus of mouse GPIHBP1 and used that antibody to coat agarose beads. We then tested the ability of soluble forms of GPIHBP1 that had been immobilized on monoclonal antibodycoated beads to bind LPL. In this assay, wild-type soluble GPI-HBP1 bound LPL avidly, but the cysteine mutants did not. Thus, our studies suggest that a structurally intact Ly6 domain (in addition to the acidic domain) is essential for LPL binding. Glycosylphosphatidylinositol-anchored high density lipoprotein-binding protein 1 (GPIHBP1)2 is an endothelial cell protein that is required for the lipolytic processing of triglyceride-rich lipoproteins in the plasma (1). In the absence of GPI-HBP1, lipolysis of plasma lipoproteins is virtually abolished, leading to severe hypertriglyceridemia (1). Expression of GPI-HBP1 in cultured cells confers the ability to bind lipoprotein lipase (LPL) (1). That finding, along with the fact that GPIHBP1 is located in endothelial cells, led Beigneux et al.(1) to hypothesize that GPIHBP1 serves as an endothelial cell platform for lipolysis.The discovery of the role of GPIHBP1 in lipolysis prompted interest in defining which portions of GPIHBP1 are important for its function (e.g. for its ability to bind LPL). Mature GPI-HBP1 is a relatively short protein with only two noteworthy structural domains (Fig. 1). First, the amino terminus of the protein contains a strongly acidic domain (amino acids 24 -48 in the mouse sequence) with a large number of aspartates and glutamates (2). This negatively charged domain is absolutely critical for binding LPL, a protein that contains two well characterized positively charged "heparin-binding domains" (3-5). Second, GPIHBP1 contains a cysteine-rich Ly6 domain (amino acids 63-135 in the mouse sequence). The function of the Ly6 domain in LPL binding is less clearly defined.The Ly6 domain is an ancient motif containing ...

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“…The precursor of human CD59 is a single peptide composed of 128 amino acids deduced from its cDNA sequence (6,7). The mature protein consists of 77 amino acids arranging in a single cysteine-rich domain composed of two antiparallel β-sheets, 5 protruding surface loops and a short helix (8)(9)(10).…”

Section: Introductionmentioning

confidence: 99%

Activity after Site-Directed Mutagenesis of CD59 on Complement-Mediated Cytolysis

et al. 2008

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Structure-Function Relationships of the Complement Regulatory Protein, CD59

Cited by 44 publications

References 43 publications

Mapping the Intermedilysin-Human CD59 Receptor Interface Reveals a Deep Correspondence with the Binding Site on CD59 for Complement Binding Proteins C8α and C9

Mapping the Intermedilysin-Human CD59 Receptor Interface Reveals a Deep Correspondence with the Binding Site on CD59 for Complement Binding Proteins C8α and C9

Highly Conserved Cysteines within the Ly6 Domain of GPIHBP1 Are Crucial for the Binding of Lipoprotein Lipase

Activity after Site-Directed Mutagenesis of CD59 on Complement-Mediated Cytolysis

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