Massachusetts Institute of Technology

Centre National De La Recherche Scientifique

Boston University

Universitè De Montpellier

Enscm

Barbara Imperiali

Melissa D Shults

Elvedin Lukovic

Dora Carrico-Moniz

Cristina Zamora

Elizabeth Sally Ward

Karen Nazaretian Allen

Eugenio Vazquez

Juan Antonio González Vera

Galen S Loving

Katherine J Franz

Juan Antonio Gonzalez-Vera

Elizabeth Ward

Glycan-binding proteins, and compositions thereof, are generally described, including methods of making and using such proteins. The proteins may include scaffolds based on easily evolvable DNA-binding proteins, with binding sites able to specifically bind to mono- or disaccharides, such as monosaccharide-binding determinants, disaccharide-binding determinants, more complex carbohydrates, etc. In certain aspects, a protein may be generated starting from a small DNA-binding protein, such as Sso7d. Such glycan-binding proteins may have numerous applications, including in enzyme-linked immunosorbent assays (ELISAs), glycan characterization, cell selection, flow cytometry, histology, imaging, arrays, affinity purification, enzyme-linked visualization, binding to a target for pharmaceutical purposes, etc.

Glycan-binding proteins and related compositions and methods

The present invention generally relates to assays of enzymes responsible for phosphoregulation including kinases (such as protein kinases, which mediate protein and peptide phosphorylation) and phosphatases (such as protein phosphatases, which mediate protein and peptide dephosphorylation). Certain aspects of the invention use lanthanide ions such as europium ions that exhibit chelation-enhanced luminescence. Phosphorylation of a peptide by a kinase may cause a complex to form between the lanthanide ion, the phosphate group, and a reporter group such as a hydroxyquinoline, which results in luminescence when in the complexed state. Thus, in certain embodiments, determination of luminescence may be indicative of kinase activity. Certain embodiments also include the use of substrates for detection of phosphatase activity, where dephosphorylation results in a loss of signal assay. Other aspects are generally related to techniques for making and using such peptides or complexes, kits involving such peptides or complexes, and the like.

Kinase and/or phosphatase sensing via hydroxyquinoline-sensitized chelates

Peptidyl sensors comprise a metal-binding peptide and one or two kinase recognition sequences with a hydroxyamino acid that can be phosphorylated in the presence of a kinase.

Sox-based kinase sensor

The present invention generally relates to environment-sensitive fluorophores, including environment-sensitive fluorophores for reporting protein/protein and peptide/protein interactions. In one aspect, the present invention is directed to compounds and salts thereof, compositions and methods useful in determining biological interactions. In some cases, the compounds of the present invention are environment-sensitive fluorophores that have spectroscopic behavior that may depend on factors such as the physicochemical properties of the surrounding environment. The compounds of the present invention can be used, in certain embodiments, to monitor ions, small molecules, and biological processes such as protein folding, protein-protein interactions and phosphorylation events.

Environmentally sensitive fluorophores

The present invention generally relates to compositions and methods for determining kinase activity. In some cases, the compositions comprise a triazole heterocycle. In some embodiments, the compositions comprise a quinoline moiety. In one aspect, the present invention is directed to compositions that undergo chelation-enhanced fluorescence (CHEF). In some cases, the compositions may have fluorescence emission spectra with peak maxima greater than 490 nm. The compositions of the present invention can be used, in certain embodiments, to detect phosphorylated substrates and biological processes such as phosphorylation events.

Kinase sensors

The present invention provides a sensor and methods for determining kinase activity.

Fluorescence assay for kinase activity

The present invention provides fluorescent compounds of formula (I) and methods monitoring protein-protein interactions.

Fluorescent probes for biological studies

The present invention provides lanthanide binding tags (LBT) that selectively complex trivalent lanthanide (Ln) ions and afford stable complexes with desirable physical properties, including at least one of fluorescence and anomalous x-ray scattering.

Growing community of inventors

Cambridge, MA, United States of America

Barbara Imperiali