Synthesis of New Phospholipids Linked to Steroid‐Hormone Derivatives Designed for Two‐Dimensional Crystallization of Proteins

Lebeau, Luc; Oudet, P; Mioskowski, Charles

doi:10.1002/hlca.19910740810

Cited by 51 publications

(42 citation statements)

References 29 publications

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“…A number of proteins have been crystallized at interfaces by targeting to surface-bound affinity ligands (1,5), by electrostatic interactions (6), and by metal coordination by polyhistidine-tagged proteins (7). The first approach requires synthesis of a lipid displaying an appropriate affinity ligand (8)(9)(10).…”

mentioning

confidence: 99%

Two-dimensional protein crystallization via metal-ion coordination by naturally occurring surface histidines.

Frey

Schief

Pack

et al. 1996

Proc. Natl. Acad. Sci. U.S.A.

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A powerful and potentially general approach to the targeting and crystallization of proteins on lipid interfaces through coordination of surface histidine residues to lipid-chelated divalent metal ions is presented. This approach, which should be applicable to the crystallization of a wide range of naturally occurring or engineered proteins, is illustrated here by the crystallization of streptavidin on a monolayer of an iminodiacetate-Cu(II) lipid spread at the air-water interface. This method allows control of the protein orientation at interfaces, which is significant for the facile production of highly ordered protein arrays and for electron density mapping in structural analysis of two-dimensional crystals. Binding of native streptavidin to the iminodiacetate-Cu lipids occurs via His-87, located on the protein surface near the biotin binding pocket. The two-dimensional streptavidin crystals show a previously undescribed microscopic shape that differs from that of crystals formed beneath biotinylated lipids.The two-dimensional (2D) crystallization of proteins on lipid monolayers has become a powerful technique with diverse applications in fields ranging from structural biology to materials science. Protein 2D crystals have been used for structure determination via electron diffraction (1, 2) and for the seeding and epitaxial growth of three-dimensional protein crystals (3, 4). Furthermore, 2D protein crystals could form the basic structure of biosensors and opto-electronic devices or act as a template for the synthesis of novel organic and inorganic materials. However, the range of soluble proteins that might be crystallized in two dimensions has been limited due to the lack of a general method for addressing proteins to lipid interfaces.A number of proteins have been crystallized at interfaces by targeting to surface-bound affinity ligands (1, 5), by electrostatic interactions (6), and by metal coordination by polyhistidine-tagged proteins (7). The first approach requires synthesis of a lipid displaying an appropriate affinity ligand (8-10). Furthermore, for many interesting proteins no convenient affinity ligand is known. Electrostatic.attraction may be successful when the protein's surface charge distribution promotes adsorption with a unique orientation at the interface, but this approach is not general, and the protein orientation is difficult to predict. Kubalek et al. (10) recently described the 2D crystallization of polyhistidine-tagged HIV-reverse transcriptase on a monolayer of chelating lipids complexed to Ni2+.Polyhistidine fusion peptides, widely used for purification of recombinant proteins by immobilized metal-affinity chromatography (IMAC) (11), provide a convenient mechanism for targeting recombinant proteins to lipid interfaces (12, 13). While this approach represents a valuable move toward more general crystallization methods, the peptide tag at the N or C terminus of the engineered protein imposes restrictions on the 2D crystal morphology and limits application to recombinant prote...

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mentioning

confidence: 99%

Two-dimensional protein crystallization via metal-ion coordination by naturally occurring surface histidines.

Frey

Schief

Pack

et al. 1996

Proc. Natl. Acad. Sci. U.S.A.

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“…The syntheses of steroid-hormone lipids 46 and 47 are shown in Scheme 9 [50]. The syntheses of 2,3-dioleoyloxypropyl ethylene glycol phosphate derivatives (33, n = 2-4) and the N-phosphatidylethanolamine derivative 1 were discussed earlier (Scheme 5).…”

Section: Steroid-hormone Derivative Lipidsmentioning

confidence: 99%

“…These lipids were to be used to facilitate crystallization of progesterone and estradiol receptors [50].…”

Section: Steroid-hormone Derivative Lipidsmentioning

confidence: 99%

Synthetic Approaches to Functionalized Lipids for Protein Monolayer Crystallizations

Hussein¹,

Ross

Landsberg³

et al. 2009

COC

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The use of lipid monolayers to bind and adsorb proteins is an attractive and increasingly important method for generating high localized concentrations of oriented proteins and protein complexes. These bound proteins can be imaged directly, or they may form 2-D crystalline arrays that are amenable to structure determination by single particle analysis or 2-D electron crystallography. 2-D crystals grown by this technique can also be used to initiate the growth of 3-D crystals for X-ray diffraction analysis. Many derivatized lipids have been prepared for use with this technique, incorporating a diverse range of ligands to enable binding to specific proteins. Synthetic lipids containing functionalized head groups that chelate Ni 2+ or Cu 2+ have also been prepared to bind and orient expressed proteins that contain His-tags. Protein-binding monolayer-forming lipids generally consist of two distinct components: (1) a branched hydrocarbon tail to confer fluidity to the monolayer and (2) a functionalized hydrophilic head group to facilitate binding of protein molecules at the air-water interface. Newer examples of these compounds also incorporate perfluorinated hydrocarbon moieties to confer detergent resistance to these lipids. This review focuses on synthetic approaches to functionalized lipids for protein monolayer crystallizations.

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“…The amine function, necessary for conjugation by dialkyl squarate chemistry, was generated at the end of the synthesis by catalytic reduction of an azide precursor (Scheme). A common approach to obtain the 8-azido-3,6-dioxaoctan-1-ol (= 2-[2-(2-azidoethoxy)ethoxy]ethanol; 4) linker relies on monosulfonylation of triethylene glycol (= 2,2'-[ethane-1,2-diylbis(oxy)]bis[ethanol]), followed by nucleophilic displacement of the leaving group with azide ion [19] [25]. The major drawback of this approach is the moderate selectivity for monoactivation of the OH groups in the starting glycol.…”

Section: Avery and Goebelmentioning

confidence: 99%

Synthesis of a Phosphorylated Disaccharide Fragment of the O‐Specific Polysaccharide of Vibrio cholerae O139, Functionalized for Conjugation

Ruttens¹,

Kòváč²

2006

Helvetica Chimica Acta

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The title disaccharide, 2-{2-{2-[(2-ethoxy-3,4-dioxocyclobut-1-en-1-yl)amino]ethoxy}ethoxy}ethyl 2-O-(3,6-dideoxy-a-L-xylo-hexopyranosyl)-b-D-galactopyranoside cyclic 4,6-(potassium phosphate) (2), was synthesized from the two isomeric linker-equipped galactose acceptors 9 and 10, obtained by phosphorylation of 2-[2-(2-azidoethoxy)ethoxy]ethyl 3-O-benzyl-b-D-galactopyranoside (8), which were glycosylated with ethyl 2,4-di-O-benzyl-3,6-dideoxy-1-thio-b-L-xylo-hexopyranoside (12; Scheme). Mainly the fully protected a-(1 ! 2)-linked products 13a and 14a were formed. Catalytic hydrogenolysis/hydrogenation effected global deprotection, thereby removing the chirality at the P-atom, and simultaneously converted the azido group in the linker to an amino group (! 15). Final treatment with diethyl squarate (= 3,4-diethoxycyclobut-3-ene-1,2-dione) gave target compound 2, amenable for conjugation to proteins.

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Synthesis of New Phospholipids Linked to Steroid‐Hormone Derivatives Designed for Two‐Dimensional Crystallization of Proteins

Cited by 51 publications

References 29 publications

Two-dimensional protein crystallization via metal-ion coordination by naturally occurring surface histidines.

Two-dimensional protein crystallization via metal-ion coordination by naturally occurring surface histidines.

Synthetic Approaches to Functionalized Lipids for Protein Monolayer Crystallizations

Synthesis of a Phosphorylated Disaccharide Fragment of the O‐Specific Polysaccharide of Vibrio cholerae O139, Functionalized for Conjugation

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