Identification of 3BP-1 in cDNA expression library by SH3 domain screening

This chapter describes a screening procedure to assay for protein–protein interactions using a biotinylated glutathione S-transferase (GST) fusion protein probe to screen cDNA expression libraries. This method is developed to allow nonradioactive probing, which has the additional advantages of a very low background signal and a low incidence of false positives. Because neither the fusion protein probe nor the protein in the expression library is denatured, this assay allows the structural components of the binding reaction to remain intact. This screening procedure offers a practical and highly efficient method of identifying protein–protein interactions. Using this procedure to detect proteins that bound to the Abl SH3 domain, five cDNA clones were isolated out of 7 million cDNA containing plaques that were screened. Of these five, three contained the identical cDNA clone termed “3BP-1,” while the other two were identical for a different clone termed “3BP-2.” On sequencing these clones, only a short stretch of approximately 40 amino acids was found where sequence similarity existed between these proteins

Identification of a ten-amino acid proline-rich SH3 binding site

The Src homology 3 (SH3) region is a small protein domain present in a very large group of proteins, including cytoskeletal elements and signaling proteins. It is believed that SH3 domains serve as modules that mediate protein-protein associations and, along with Src homology 2 (SH2) domains, regulate cytoplasmic signaling. The SH3 binding sites of two SH3 binding proteins were localized to a nine- or ten-amino acid stretch very rich in proline residues. Similar SH3 binding motifs exist in the formins, proteins that function in pattern formation in embryonic limbs of the mouse, and one subtype of the muscarinic acetylcholine receptor. Identification of the SH3 binding site provides a basis for understanding the interaction between the SH3 domains and their targets

A Nuclear Tyrosine Kinase Becomes a Cytoplasmic Oncogene

It is a great pleasure to be here at this celebration. My whole scientific career has, of course, been shaped by the discovery in 1953 of the structure of DNA. At that time I was in high school, and hardly aware of it. But I do remember an incident in the late 1950s when, as a college student, I had the rare opportunity to drive Jim Watson from Cold Spring Harbor to a nearby airport on Long Island. At that time he said to me “There’s a virus that’s just been discovered that has only a small amount of DNA in it.” It had to be SV40 or polyoma virus. “That such a virus is able to cause cancer means that a very small amount of genetic information is all that’s required to cause cancer and we should be able to decipher that very quickly,” he observed. Well, as usual, his insight was remarkable, because those viruses have played a central role in developing the notions of oncogenes; his time line, however, was a little short

Il modello organizzativo dipartimentale nel Servizio sanitario nazionale: risultati dell\u2019indagine nazionale

During the \u201990s the Italian national health service has undergone important reforms. These introduced new organizational and managerial models within hospitals. The main aim of the new organizational model, named clinical directorate, is the coordination of specialized care services. Moreover clinical directorates have been recognised as the context in which clinical governance tools can be developed. The study presents methods and results of a national wide survey of the introduction of clinical directorates in Italian public hospitals and local health units performed in 2005. The survey was conducted by Italian Ministry of Health in collaboration with the Catholic University of the Sacred Heart, Rome