ABSTRACT
A simple and rapid strategy is described to screen protein fractions for defined enzymatic activity. A protein fraction from a porcine kidney extract was immobilized by covalent coupling to activated affinity beads. The immobilized proteins were incubated with probes specific for different enzyme activities. The reaction products were analyzed by matrix-assisted laser desorption/ionization (MALDI)-mass spectrometry. The MALDI spectra indicate the presence of 5'-nucleotidase, phosphatase, kinase, glutathione reductase, and renin activities in the kidney protein extract. Furthermore, the method can be used to screen for inhibitors of enzymatic reactions. The method is adaptable to high-throughput sample handling and automated mass spectrometric analysis and therefore suited for functional genomics.
Subject(s)
Enzymes/analysis , Kidney/enzymology , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods , Animals , Electrophoresis, Polyacrylamide Gel , Enzyme Inhibitors/analysis , Microspheres , Substrate Specificity , Swine , Time FactorsABSTRACT
A biomechanical test system has been developed and validated to conduct controlled uniaxial impact experiments of head and facial trauma. The design reduces off-axis accelerations which are not in the direction of impact and allows accurate positioning of test specimens. Impact forces, displacement histories, impulses at impact and spectral responses are compared to free-fall test results at contact velocities representative of facial injuries (2.5, 3.1 and 3.8 m s-1). Models based on the experimental results are developed to reveal stiffness and inertial properties of impact for use in the design of biomechanically protective steering wheels, air bags and other potential impact structures. The results indicate that the system provides a flexible yet controllable method for positioning and testing impact structures reliably.
