Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.4.21.4 (trypsin)
 42,187 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Two high-affinity monoclonal antibodies (ROS-1, ROS-2) have been produced to the rod outer segment phosphodiesterase (ROS PDE). These antibodies bind at different antigenic determinants. ROS-2 absorbs a subset of the total PDE activity from a detergent-solubilized retina extract, whereas ROS-1 adsorbs nearly all of the PDE activity. DEAE-cellulose chromatography separates two peaks of activity from a hypotonic extract of rod outer segments. Peak I activity is adsorbed only by ROS-1, whereas peak II activity is adsorbed by both ROS-1 and ROS-2. Both peaks of activity are activated by histone H2B and limited trypsin digestion, and both peaks of activity contain a heat-stable, trypsin-sensitive inhibitor. When analyzed by SDS gel electrophoresis, ROS-1 adsorbed a single peptide from peak I, which comigrated with phosphorylase B, whereas ROS-1 adsorbed two slightly faster migrating peptides from peak II. Histone H2B activated at least 80% of the PDE activity bound to ROS-2 but was less effective in activating the PDE bound to ROS-1. ROS-1 but not ROS-2 was an effective inhibitor of PDE activity, suggesting that ROS-1 may be a specific probe to study the effects of ROS PDE on the light response.
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PMID:Immunotitration analysis of the rod outer segment phosphodiesterase. 620 41

The purpose of this study is to determine the feasibility of the direct matrix-assisted laser desorption/ionization (MALDI) identification of proteins in fixed T47D breast cancer cells and murine brain tissues. The ability to identify proteins from cells and tissue may lead to biomarkers that effectively predict the onset of defined disease states, and their dynamic behavior could be an important hint for drug target discoveries. Direct tissue application of trypsin allows protein identification in cells and tissues, while maintaining spatial integrity and intracellular organization. Using a chemical printer, matrix was co-registered on trypsinized human T47D breast cancer cells and cryo-preserved sections of murine brain tissue, followed by MALDI post-source decay (PSD) or MALDI collision-induced dissociation (CID), respectively. Mass-to-charge (m/z) data from the cells and brain tissues were processed using Mascot software interrogation of the National Center for Biotechnology Information (NCBI) database. Histone H2B was identified from cultured T47D human breast cancer cells. Tubulin beta2 was identified from mouse brain cortex following an induced stroke. These results suggest that MALDI PSD/CID, combined with bioinformatics, can be used for the direct identification of proteins from cells and tissues. Refinements in preparation techniques may improve this approach to provide a tool for quantitative proteomics and clinical analysis.
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PMID:Direct identification of proteins from T47D cells and murine brain tissue by matrix-assisted laser desorption/ionization post-source decay/collision-induced dissociation. 1721 66