Attaching histidine-tagged peptides and proteins to lipid-based carriers through use of metal-ion-chelating lipids

Chikh, Ghania; Li, Wai Ming; Schutze-Redelmeier, Marie-Paule; Meunier, Jean‐Claude; Bally, Marcel B.

doi:10.1016/s0005-2736(02)00618-1

Cited by 59 publications

(53 citation statements)

References 27 publications

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“…One promising approach to noncovalent antigen conjugation involves metal chelation, in which polyhistidine-tagged proteins are attached to nitrilotriacetic acid (NTA)-containing liposomes or microparticles with micromolar affinity (9,13,44). Since NTA-Ni(II)-His binding is site specific, the physical orientation of the antigen on the particulate surface can be controlled.…”

mentioning

confidence: 99%

“…Liposomes were an appropriate delivery system for this study because they deliver associated antigens efficiently to antigen-presenting cells (12) and stimulate potent immune responses when adjuvanted with monophosphoryl lipid A (MPL) (5), and lipidanchored NTA molecules can be readily incorporated into the formulation (9,22). Ovalbumin was selected because it is widely used as a model antigen for assessment of humoral and cellular responses (7,50).…”

mentioning

confidence: 99%

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Antibody Response to Polyhistidine-Tagged Peptide and Protein Antigens Attached to Liposomes via Lipid-Linked Nitrilotriacetic Acid in Mice

Watson

Platt

Cao

et al. 2011

Clin Vaccine Immunol

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confidence: 99%

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confidence: 99%

Antibody Response to Polyhistidine-Tagged Peptide and Protein Antigens Attached to Liposomes via Lipid-Linked Nitrilotriacetic Acid in Mice

Watson

Platt

Cao

et al. 2011

Clin Vaccine Immunol

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“…They have also been used recently to attach oligohistidine-tagged peptides to liposomes, as a delivery method for peptides to targeted cells (21); however, they have rarely been used to attach proteins to membranes for functional studies (22). We reasoned that incorporating a hexahistidine tag at the C-terminus of sTF would allow it to bind to chelated nickel ions at the membrane surface and become an effective protein cofactor for fVIIa (but at much lower surface densities than when preparing two-dimensional protein crystals!).…”

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confidence: 99%

Restoring Full Biological Activity to the Isolated Ectodomain of an Integral Membrane Protein

Waters

Morrissey

2006

Biochemistry

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Integral membrane proteins, which include many cellular effector proteins and drug targets, can be difficult to produce, purify and manipulate. Although the isolated ectodomains of many membrane proteins can be expressed as water soluble proteins, biological activity is frequently lost when these proteins are released from the membrane surface. An example is tissue factor, the integral membrane protein that triggers the blood clotting cascade and for which membrane anchoring is essential: Its isolated ectodomain (soluble tissue factor) can be expressed with high yield in bacteria but is orders of magnitude less active than the intact, membrane-anchored protein. We now report full restoration of biological activity to the isolated tissue factor ectodomain by engineering a hexahistidine tag onto its C-terminus and using it in combination with membrane bilayers containing nickel-chelating lipids. When soluble tissue factor was tethered to the membrane surface via such metal-chelating lipids, it bound factor VIIa with the same high affinity as wild-type tissue factor, and the resulting factor VIIatissue factor complexes supported factor X activation and factor VII autoactivation with essentially wild-type enzyme kinetic constants. Furthermore, when such bilayers were immobilized onto solid supports they efficiently captured histidine-tagged soluble tissue factor directly from crude culture supernatants -with full biological activity -obviating the need for purification or laborious membrane reconstitution procedures. This strategy is rapid, efficient, scalable, automatable, and should be applicable to other integral membrane proteins, especially those with a single transmembrane domain. Applications include high-throughput screening of mutants or drugs, flow reactors, clinical assays and point-of-care instrumentation.Integral membrane proteins represent an estimated 20-30% of the proteins in many genomes and are a major class of current and future drug targets (1-3). They are, however, more difficult to express at high yield than are soluble proteins, and they require detergents to extract them from membranes and keep them soluble during purification, which may diminish their stability and activity. Although effective procedures have been developed to reconstitute purified integral membrane proteins into phospholipid vesicles, most are laborious, time-consuming, and expensive (4).The isolated ectodomains of some integral membrane proteins can be expressed with very high yield in recombinant systems and are generally far easier to handle than are their membraneanchored counterparts. However, the soluble ectodomains of some membrane proteins have little or no biological activity. An example is tissue factor (TF 1 ) the integral membrane protein responsible for triggering the blood clotting cascade in normal hemostasis and many thrombotic † Supported by grant R01 HL47014 from the National Heart Lung & Blood Institute. E.K.W. was supported in part by NIH training grant T32 GM007283. * To whom correspondence shou...

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“…Application of Ni-NTA-DOGS liposomes for the construction of experimental vaccine against sytemic Candida infection based on Candida Heat shock protein 90 kDa (rHSP90-HisTag) showed good stability in serum as well as strong immune response against recobinant rHSP90-HisTag antigen in mice (Masek et al, 2011b). In vivo activity (immunogenicity) was also demonstrated for antigens associated with ISCOM particles via metallochelating lipid dipalmitoyliminodiacetic acid (Malliaros et al, 2004) and a peptide antigen associated with liposomes via Ni-NTA-DOGS (Chikh et al, 2002). Generally, metal ions, physico-chemical character of the metallochelating lipids and their surface density on the particles belong to the factors that could be optimized to get a required in vivo stability and, therefore, a strong immune response.…”

Section: Metallochelating Bond and Its Application For Construction Omentioning

confidence: 99%

“…Both reversible character and high affinity of the metallochelating bonds are very useful for the preparation of various self-assembling supramolecular structures useful for the construction of experimental vaccines (Chikh et al, 2002;Malliaros et al, 2004;Masek et al, 2011a;Masek et al, 2011b). As an example of synthetic liposome-based recombinant vaccine we can use metallochelating liposomes and recombinant antigen rOspc-6HisTag derived from the pathogen Borrelia burgdorferi.…”

Section: Metallochelation Liposomes For Construction Of Experimental mentioning

confidence: 99%

Application of Liposomes for Construction of Vaccines

Tuřánek¹,

Mašek²,

Raška³

et al. 2012

Biomedical Science, Engineering and Technology

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Attaching histidine-tagged peptides and proteins to lipid-based carriers through use of metal-ion-chelating lipids

Cited by 59 publications

References 27 publications

Antibody Response to Polyhistidine-Tagged Peptide and Protein Antigens Attached to Liposomes via Lipid-Linked Nitrilotriacetic Acid in Mice

Antibody Response to Polyhistidine-Tagged Peptide and Protein Antigens Attached to Liposomes via Lipid-Linked Nitrilotriacetic Acid in Mice

Restoring Full Biological Activity to the Isolated Ectodomain of an Integral Membrane Protein

Application of Liposomes for Construction of Vaccines

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