An accurate method for the quantitation of Fmoc-derivatized solid phase supports

Gude, Markus; Ryf, Jeannine; White, Peter

doi:10.1007/bf02538384

Cited by 100 publications

(127 citation statements)

References 3 publications

Supporting

Mentioning

125

Contrasting

Unclassified

Order By: Relevance

“…Preparation of Lipid-Peptide Complexes: For CD spectroscopy, solutions of large unilamellar vesicles (LUVs) were prepared from a phosphate buffer solution (1 mm NaH 2 PO 4 /Na 2 HPO 4 , pH 7.0) following a method from MacDonald et al [52] For preparation of the lipid films the DLPC dissolved in CHCl 3 (10 mg mL…”

Section: Methodsmentioning

confidence: 99%

A Novel Heavy‐Atom Label for Side‐Specific Peptide Iodination: Synthesis, Membrane Incorporation and X‐ray Reflectivity

et al. 2009

View full text Add to dashboard Cite

Structural parameters, such as conformation, orientation and penetration depth of membrane-bound peptides and proteins that may function as channels, pores or biocatalysts, are of persistent interest and have to be probed in the native fluid state of a membrane. X-ray scattering in combination with heavy-atom labeling is a powerful and highly appropriate method to reveal the position of a certain amino acid residue within a lipid bilayer with respect to the membrane normal axis up to a resolution of several Angstrøm. Herein, we report the synthesis of a new iodine-labeled amino acid building block. This building block is intended for peptide incorporation to provide high intensities for electron density difference analysis of X-ray reflectivity data and improve the labeling potential for the lipid bilayer head-group and water region. The novel building block as well as the commercially available non-iodinated analogue, required for X-ray scattering, was implemented in a transmembrane peptide motif via manual solid-phase peptide synthesis (SPPS) following the fluorenylmethyloxycarbonyl (Fmoc)-strategy. The derived peptides were reconstituted in lipid vesicles as well as in highly aligned multilamellar lipid stacks and investigated via circular dichroism (CD) and X-ray reflectivity. Thereby, it has been revealed that the bulky iodine probe neither causes conformational change of the peptide structure nor lamellar disordering of the membrane complexes.

show abstract

Section: Methodsmentioning

confidence: 99%

A Novel Heavy‐Atom Label for Side‐Specific Peptide Iodination: Synthesis, Membrane Incorporation and X‐ray Reflectivity

et al. 2009

View full text Add to dashboard Cite

show abstract

“…This result can be ascribed to the loading of the Fmoc-Gly-DFPE resin 3. Indeed, Fmoc quantification [11] gave a loading of 0.77 versus 0.92 mmol g À1 for the commercially available DFPE resin. Despite monitoring by colorimetric tests, the reductive amination/acylation reactions were not complete (83 % yield based on the loading of the DFPE resin).…”

Section: Wwwchemeurjorgmentioning

confidence: 99%

Solid‐Phase Organic Tagging Resins for Labeling Biomolecules by 1,3‐Dipolar Cycloaddition: Application to the Synthesis of a Fluorescent Non‐Peptidic Vasopressin Receptor Ligand

Bron¹,

Riché²,

Loison³

et al. 2008

Chemistry A European J

View full text Add to dashboard Cite

Two novel solid-phase organic tagging (SPOrT) resins were synthesized to facilitate the labeling of peptides and small organic compounds with a fluorescent probe. Both resins were obtained from the commercially available backbone amide linker (BAL) resin. Following the solid-phase synthesis of model compounds, a tripeptide and benzazepine, the fluorescent probe derived from Lissamine Rhodamine B was incorporated through CuI-catalyzed 1,3-dipolar cycloaddition. Final cleavage in acidic media enabled access to both types of molecules in good yield with high purity. The SPOrT resin was successfully applied to the preparation of the first non-peptidic fluorescent compound with a nanomolar affinity for the human vasopressin V2 receptor (V2R) subtype. This molecule will find application in binding assays that use polarization or fluorescence resonance energy-transfer (FRET) techniques. The SPOrT resins are also well suited for other tags and the parallel synthesis of a fluorescently tagged library for protein screening.

show abstract

“…[70,105] However, other groups reported different molar extinction coefficients [106] and fluctuating DG values which can most likely be attributed to the equilibrium of PDFA and dibenzofulvene/piperidine. [107] Therefore, this method is not considered to provide absolute quantities, but values which can be compared to results obtained in previous works in the same manner. [105,108] To determine the DG of irregularly shaped silicon wafer pieces, the surface area A was calculated from the weight of the wafer piece.…”

Section: Smartspecs Plusmentioning

confidence: 99%

“…Furthermore, the equilibrium between PDFA and dibenzofulvene/piperidine is assumed to affect the experimental results. [107] Hence, all coupling efficiencies calculated on the basis of the PDFA method for the 10:90-PEGMA-co-PMMA-NH2…”

Section: Iii22 Coupling Of Cleavable Linkersmentioning

confidence: 99%

Purification of Peptides in High-Complexity Arrays

Schirwitz

2013

Springer Theses

View full text Add to dashboard Cite

Compared to the human genome, which is formed by approximately 25,000 genes, the human proteome comprises over a million functional proteins and thus represents a much higher diversity. Although genetic research provides valuable information on the proteins which can be translated, a great task in the future will be the exploration of the entire protein interaction network. Understanding protein interactions will greatly help scientists to identify the mechanisms behind fatal diseases such as cancer, AIDS, and tuberculosis and, hopefully, provide new

show abstract

An accurate method for the quantitation of Fmoc-derivatized solid phase supports

Cited by 100 publications

References 3 publications

A Novel Heavy‐Atom Label for Side‐Specific Peptide Iodination: Synthesis, Membrane Incorporation and X‐ray Reflectivity

A Novel Heavy‐Atom Label for Side‐Specific Peptide Iodination: Synthesis, Membrane Incorporation and X‐ray Reflectivity

Solid‐Phase Organic Tagging Resins for Labeling Biomolecules by 1,3‐Dipolar Cycloaddition: Application to the Synthesis of a Fluorescent Non‐Peptidic Vasopressin Receptor Ligand

Purification of Peptides in High-Complexity Arrays

Contact Info

Product

Resources

About