title: Elimination of false negative results in the two-hybrid system in the phagocyte NADPH oxidase.
abstract: The yeast two-hybrid system is finding increased use in the study of interactions between proteins. In this method, two polypeptides are expressed in yeast as fusion proteins to a transcriptional activator DNA-binding domain (bd) and activating domain (ad), respectively. Interaction between the two polypeptides reconstitutes function of a transactivator which controls expression of reporters. The phagocyte NADPH oxidase is a complex of membrane cytochrome b558 (comprised of subunits p22-phox and gp91-phox) and three cytosol proteins (p47-phox, p67-phox, and p21rac) that translocate to membrane and bind to cytochrome b558. This is the first report to demonstrate that two of cytosolic components of cytochrome b558, p47-phox binding to p67-phox each other. We encountered several methodological problems in the two-hybrid system which are the focus of this report.
authors.affiliation: Graduate Institute of Biochemistry, Kaohsiung Medical College, Taiwan, Republic of China.
authors.forename: Y R
authors.initials: YR
authors.lastname: Hong
authors.forename: L
authors.initials: L
authors.lastname: Han
authors.forename: J
authors.initials: J
authors.lastname: Huang
authors.forename: M E
authors.initials: ME
authors.lastname: Kleinberg
mesh_terms: D005188:False Negative Reactions; D006801:Humans; D009247:NADH, NADPH Oxidoreductases / Q000032:analysis*; D019255:NADPH Oxidases; D010586:Phagocytes / Q000201:enzymology*; D012441:Saccharomyces cerevisiae / Q000235:genetics*
chemical_list: D009247:NADH, NADPH Oxidoreductases; D019255:NADPH Oxidases