Polyglycerol-bound phosphotriesterase enzyme model complexes for detection and hydrolysis of phosphorus species in aqueous solution

Mangalum, Anshuman; Smith, Rhett C.

doi:10.1016/j.tet.2009.03.051

Cited by 10 publications

(7 citation statements)

References 38 publications

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“…A similar approach has been reported by Scrimin et al, who had used Tentagel resin as support (pendant NH 2 groups) [22]. Attempts to hydrolyze the ester group of CO 2 EtH 3 L1 with NaOH in ethanolic solution were unsuccessful although this had been described in the literature for the similar ligand methyl 3,5-bis [9-bis(2-pyridylmethyl)amino)methyl]-hydroxybenzoate [23]. Initially it was thought that the alcohol arms were causing the problems during the alkaline hydrolysis.…”

Section: Early Attempts To Immobilize/polymerize Model Complexesmentioning

confidence: 80%

See 1 more Smart Citation

Immobilization of GpdQ and Related Models for Bioremedial Applications

Daumann

2014

Spectroscopic and Mechanistic Studies of Dinuclear Metallohydrolases and Their Biomimetic Complexes

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Section: Early Attempts To Immobilize/polymerize Model Complexesmentioning

confidence: 80%

“…This resulted in only very low yields of the desired product. After a close examination of the supporting information it was clear that this type of reaction was not very clean and thus other methods had to be evaluated [23].…”

Section: Early Attempts To Immobilize/polymerize Model Complexesmentioning

confidence: 99%

Immobilization of GpdQ and Related Models for Bioremedial Applications

Daumann

2014

Spectroscopic and Mechanistic Studies of Dinuclear Metallohydrolases and Their Biomimetic Complexes

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“…Phosphorylation is the most commonly occurring posttranslational modification [41] and a small molecule binder for phosphate was linked to the polypeptides in the search for binder molecules for groups of phosphorylated proteins, possibly all [42]. The small molecule ligand complexes two Zn 2+ ions and binds phosphate ions with dissociation constants reported to be in the range from 28 to 640 μmol L −1 [43]. The small molecule ligand was conjugated to the side chain of a lysine residue in eight of the polypeptides without connecting linker, because the objective was to bind phosphoserines and phosphothreonines that do not extend far from the protein surface (Fig.…”

Section: Polypeptide Conjugate Binder Moleculesmentioning

confidence: 99%

Crossing borders to bind proteins—a new concept in protein recognition based on the conjugation of small organic molecules or short peptides to polypeptides from a designed set

Baltzer

2011

Anal Bioanal Chem

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A new concept for protein recognition and binding is highlighted. The conjugation of small organic molecules or short peptides to polypeptides from a designed set provides binder molecules that bind proteins with high affinities, and with selectivities that are equal to those of antibodies. The small organic molecules or peptides need to bind the protein targets but only with modest affinities and selectivities, because conjugation to the polypeptides results in molecules with dramatically improved binder performance. The polypeptides are selected from a set of only sixteen sequences designed to bind, in principle, any protein. The small number of polypeptides used to prepare high-affinity binders contrasts sharply with the huge libraries used in binder technologies based on selection or immunization. Also, unlike antibodies and engineered proteins, the polypeptides have unordered three-dimensional structures and adapt to the proteins to which they bind. Binder molecules for the C-reactive protein, human carbonic anhydrase II, acetylcholine esterase, thymidine kinase 1, phosphorylated proteins, the D-dimer, and a number of antibodies are used as examples to demonstrate that affinities are achieved that are higher than those of the small molecules or peptides by as much as four orders of magnitude. Evaluation by pull-down experiments and ELISA-based tests in human serum show selectivities to be equal to those of antibodies. Small organic molecules and peptides are readily available from pools of endogenous ligands, enzyme substrates, inhibitors or products, from screened small molecule libraries, from phage display, and from mRNA display. The technology is an alternative to established binder concepts for applications in drug development, diagnostics, medical imaging, and protein separation.

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“…25 The synthesis route was optimized, see supporting information, and the target compound produced in very good yield compared to that reported in the literature. 17 The binder molecules were formed in a reaction between 5 and the lysine side chain of the peptide attached to the solid support using PyBOP (benzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate) to ensure a high yield. The binder was cleaved from the resin and purified by HPLC.…”

Section: Binder Design and Synthesismentioning

confidence: 99%

“…Here, we have addressed the problem of recognition and binding of phosphorylated proteins, and in a proof-of-principle demonstration applied the binder concept to the capturing of the model proteins a-casein, b-casein and ovalbumin. 16,17 Both caseins carry multiple phosphate groups that may enhance the probability for binding and thus their avidities. The binder molecules have been evaluated with regards to affinity, binding mechanism and selectivity in media of variable complexity.…”

Section: Introductionmentioning

confidence: 99%

The molecular recognition of phosphorylated proteins by designed polypeptides conjugated to a small molecule that binds phosphate

Ślósarczyk

Baltzer

2011

Org. Biomol. Chem.

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The conjugation of polypeptides from a designed set to the small molecule ligand 3,5-bis[[bis(2-pyridylmethyl)amino]methyl]benzoic acid, which in the presence of Zn(2+) ions binds inorganic phosphate, has been shown to provide a polypeptide conjugate that binds α-casein, a multiply phosphorylated protein, with a dissociation constant K(D) of 17 nM. The measured affinity is more than three orders of magnitude higher than that of the small molecule ligand for phosphate and the binding of 500 nM of α-casein was not inhibited by 10 mM phosphate buffer, providing a 2000-fold excess of phosphate ion over protein. The selectivity for phosphoproteins was demonstrated by extraction of α-casein from solutions of various complexity, including milk and human serum spiked with α-casein. In addition to α-casein, β-casein was also recognized but not ovoalbumin. Conjugation of a polypeptide to the zinc chelating ligand was therefore shown to give rise to dramatically increased affinity and also increased selectivity. A set of polypeptide conjugates is expected to be able to capture a large number of phosphorylated proteins, perhaps all, and in combination with electrophoresis or mass spectrometry become a powerful tool for the monitoring of phosphorylation levels. The presented binder can easily be attached to various types of surfaces; here demonstrated for the case of polystyrene particles. The example of phosphoproteins was selected since posttranslational phosphorylation is of fundamental importance in cell biology due to its role in signaling and therefore of great interest in drug development. The reported concept for binder development is, however, quite general and high-affinity binders can conveniently be developed for a variety of proteins including those with posttranslational modifications for which small molecule recognition elements are available.

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Polyglycerol-bound phosphotriesterase enzyme model complexes for detection and hydrolysis of phosphorus species in aqueous solution

Cited by 10 publications

References 38 publications

Immobilization of GpdQ and Related Models for Bioremedial Applications

Immobilization of GpdQ and Related Models for Bioremedial Applications

Crossing borders to bind proteins—a new concept in protein recognition based on the conjugation of small organic molecules or short peptides to polypeptides from a designed set

The molecular recognition of phosphorylated proteins by designed polypeptides conjugated to a small molecule that binds phosphate

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