Two-dimensional gel pattern of proteins synthesized de novo in lymphoid organs of the mouse.

Abstract

Abstract Mice were injected intravenously with 1 mCi of [35S]methionine, and serum and tissue were sampled 4 h later. Serum and lymphoid and non-lymphoid tissues had all incorporated enough labeled methionine to allow radiofluorographic detection on two-dimensional gels of proteins synthesized de novo and, by comparing radiofluorographs, we could distinguish spots peculiar to a given tissue from others more ubiquitous. We selected a protein of 17-kDa apparent molecular mass and pl about 4 to demonstrate tissue specificity. All patterns obtained for thymus samples yielded this spot; in all other immunologically relevant sites it was missing or weak. It also was not detected on gels previously obtained from lymphocyte subpopulations biosynthetically labeled in vitro. The labeling method described here will be especially helpful for characterizing cell populations that cannot be radiolabeled under cell-culture conditions. In contrast to detection methods that detect all proteins of a sample, in vivo labeling allows specific recognition of de novo synthesized proteins. Therefore it will facilitate comparison and detection of proteins produced by an animal in response to a given treatment.