Johns Hopkins University

University System of Maryland

Min Li

Heng Zhu

Sojin Shikano

Jef D Boeke

Haiyan Sun

Prashant Desai

Shaohui Hu

Shunyou Long

John J Laterra

Amy M Fulton

Han Sun

Provided herein are recombinant virion arrays comprising human membrane bound proteins that retain their native conformations and/or interactions, recombinant HSV-1 virions, and methods of use including high-content, high-throughput assays for screening for ligands and/or drugs that bind human membrane bound proteins, diagnostic assays, proteomic assays, and biosensor assays. Also provided are recombinant HSV-1 virions comprising an envelope comprising a plurality of heterologous membrane bound proteins that retain their native conformations and/or interactions as well as recombinant HSV-1 bacterial artificial chromosome (BAC) clones encoding heterologous membrane polypeptides.

Virion display array for profiling functions and interactions of human membrane proteins

Provided herein are systems and methods for storage, retrieval and sampling of materials.

System and methods for sampling materials

Membrane proteins represent &#x2dc;30% of the proteome of both prokaryotes and eukaryotes. Unique to cell surface receptors is their biogenesis pathway, which involves vesicular trafficking from the endoplasmic reticulum through the Golgi apparatus and to the cell surface. Increasing evidence suggests specific regulation of biogenesis for different membrane receptors, hence affecting their surface expression. A pulse-chase assay can be used to monitor function recovery after chemobleaching (FRAC) to probe the transit time of the cell surface receptors to reach the cell surface. This method distinguishes molecular density from functional density. The ability of the reported method to access the biogenesis pathways in a high-throughput manner facilitates the identification and evaluation of molecules affecting receptor trafficking.

Method to assay test substances

The invention provides amino acid sequence motifs (e.g., biomolecule partition motifs) that can direct targeting of intracellular polypeptides to and through membranes, including cell surface membranes.

Biomolecule partition motifs and uses thereof

Viruses expressing ligands on their surfaces are used as a detection means for the related polypeptide which binds the ligand. Multiple copies of the ligand can be expressed on the viral surface. These viruses may be used to detect polypeptides, cells, receptors and channel proteins.

Methods of detection utilizing modified bacteriophage

A polypeptide consisting essentially of the NAB and linking region of an .alpha.-subunit of Shaker-like potassium ion channel which binds to a core region of a .beta.-subunit of said Shaker-like potassium ion channel. A related polypeptide is also provided and consists essentially of the core region of a .beta.-subunit of a Shaker-like potassium ion channel which binds to the NAB and linking region of an .alpha.-subunit of said Shaker-like channel. Nucleic acid sequences which encode these polypeptides, vectors containing those sequences, expression systems, hosts cells containing the aforesaid polypeptides, and pharmaceutical formulations of the peptides are also provided. Other aspects of the invention include methods of modulating the flow of potassium ions through a cell membrane surrounding a cytoplasm by introducing either of the aforesaid polypeptides or exogenous Kv.beta.2 protein into the cytoplasm of the cell, methods of detecting a molecule that binds to either of the aforesaid polypeptides, and an improved yeast two-hybrid system which utilizes the aforesaid polypeptides.

Growing community of inventors

Lutherville, MD, United States of America

Min Li