A substoichiometric radioisotope dilution study using210Pb

A radioisotope dilution procedure has been developed which permits the determination of lead over a concentration range of 0.1 to 10 ppm. Samples in an ammoniacal cyanide/sulfite medium are labelled with²¹⁰Pb, followed by addition of substoichiometric amount of dithizone. The Pb/HDz/₂ formed is extracted into CH₂Cl₂, and aliquot portions are taken for liquid scintillation counting. Count rate is plotted vs. the reciprocal of the lead concentration, yielding a straight line. An average error of ±3.5% is estimated, based on a linear least squares fit.     

LASER CROSS-LINKING OF PROTEINS TO NUCLEIC ACIDS: PHOTODEGRADATION AND ALTERNATIVE PHOTOPRODUCTS OF THE BACTERIOPHAGE T4 GENE 32 PROTEIN

Pulsed laser cross-linking provides a means of introducing a covalent bond between proteins and the nucleic acids to which they are bound. This rapid cross-linking effectively traps the equilibrium that exists at the moment of irradiation and thus allows examination of the protein-nucleic acid interactions that existed. Laser irradiation may also induce photodestruction of protein and we have used the bacteriophage T4 gene 32 protein to investigate this phenomenon. Our results show that both nonspecific and specific photoproducts can occur, specifically at wavelengths where the peptide backbone of proteins is known to absorb. These results demonstrate that nonspecific photodegradation can be correlated with the formation of a specific photodegradation product. The formation of this product was monitored to show that product yield is nonlinearly dependent on laser power and wavelength. We have also investigated an unexpected photoproduct whose formation is dependent on the length of the polynucleotide to which the gene 32 protein binds and that further demonstrates the complexities of analyzing protein-nucleic acid interactions through the use of UV laser cross-linking. These data provide essential information for the establishment of appropriate conditions for future studies that use UV cross-linking of protein-nucleic acid complexes.