Development of a 1-microl scale assay for mitogen-activated kinase kinase 7 using 2-D fluorescence intensity distribution analysis anisotropy.

This paper describes the development of a robust, miniaturizable, and quantitative fluorescence-based assay for mitogen-activated protein kinase kinase 7 (MKK7). As a first step, the basic steady-state kinetics of the MKK7-catalyzed phosphorylation of c-Jun N-terminal kinases (JNKs) 1, 2, and 3 were defined using standard radiometric methods. Subsequently, the authors found that in addition to the holo JNKs, a series of novel small peptides (based on the region around the JNK phosphorylation site) are also substrates, provided that these were prephosphorylated on the Y residue of the TPY motif.

The application of confocal microscopy to the study of liposome adsorption onto bacterial biofilms.

Confocal laser scanning microscopy has been used to visualise the adsorption of fluorescently labelled liposomes on immobilised biofilms of the bacterium Staphylococcus aureus. The liposomes were prepared with a wide range of compositions with phosphatidylcholines as the predominant lipids using the extrusion technique. They had weight average diameters of 125 +/- 5 nm and were prepared with encapsulated carboxyfluorescein.