A TRANSMEMBRANE TRAP FOR ISOLATING MEMBRANE BOUND PROTEINS

摘要

The present invention relates to methods of identifying the transmembrane domain of membrane-spanning proteins. The membrane-spanning proteins can be Group I receptors, Group II receptors and receptors with multiple transmembrane domains. The methods relate to the use of a death domain-lacki ng protein, modified to replace its transmembrane domain with a candidate nucle ic acid sequence. The absence of apoptosis of a host cell expressing the modifi ed death domain-lacking protein and a death domain-containing protein following exposure to an apoptosis-inducing ligand denotes the presence of a candidate nucleic acid sequence encoding a transmembrane domain of a membrane-spanning receptor. Vectors and host cells producing the modified death domain-lacking protein are also provided. The invention further relates to the identificati on of the membrane-spanning proteins containing the transmembrane domains identified by the methods of the invention and target molecules for the membrane-spanning proteins. The invention also includes methods of identifyi ng substances which alter the interaction of the membrane-spanning proteins and target molecules identified by methods of the invention.