Metabolic labeling and immunoprecipitation of yeast proteins.
The proteins of Saccharomyces cervsiae can be metabolically labeled, as described here, with (35)methionine and (35)cysteine or a hydrolysate of E. coli labeled with (35)O4(2-). After the labeling, protocols are provided for the mechanical disruption of the yeast cells or conversion to spheroplasts, with subsequent lysis before immunoprecipitation of the proteins.
Graham
T R
TR
Vanderbilt University, Nashville, Tennessee, USA.
D003545:Cysteine / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D047468:Immunoprecipitation / Q000379:methods*; D007202:Indicators and Reagents; D007553:Isotope Labeling / Q000379:methods; D008715:Methionine / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000166:cytology; D029701:Saccharomyces cerevisiae Proteins / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D013104:Spheroplasts / Q000737:chemistry; D013431:Sulfates / Q000378:metabolism; D013462:Sulfur Radioisotopes / Q000032:analysis / Q000378:metabolism
D007202:Indicators and Reagents; D029701:Saccharomyces cerevisiae Proteins; D013431:Sulfates; D013462:Sulfur Radioisotopes; D008715:Methionine; D003545:Cysteine