Application of high-performance liquid chromatographic methods in the monitoring of enzyme-targeted chemotherapy

Lipid rafts are cholesterol-sphingolipid-rich microdomains that function as platforms for membrane trafficking and signal transduction. Caveolae are specialized lipid raft domains that contain the structural proteins known as the caveolins. Connexins are a family of transmembrane proteins that self-associate to form cell-cell connections known as gap junctions and that are linked to cytosolic proteins, forming a protein complex or Nexus. To determine the extent to which these intracellular compartments intersect, we have systematically evaluated whether connexins are associated with lipid rafts and caveolin-1. We show that connexin 43 (Cx43) colocalizes, cofractionates, and coimmunoprecipitates with caveolin-1. A mutational analysis of Cx43 reveals that the hypothesized PDZ- and presumptive SH2/SH3-binding domains within the Cx43 carboxyl terminus are not required for this targeting event or for its stable interaction with caveolin-1. Furthermore, Cx43 appears to interact with two distinct caveolin-1 domains, i.e., the caveolin-scaffolding domain (residues 82-101) and the C-terminal domain (135-178). We also show that other connexins (Cx32, Cx36, and Cx46) are targeted to lipid rafts, while Cx26 and Cx50 are specifically excluded from these membrane microdomains. Interestingly, recombinant coexpression of Cx26 with caveolin-1 recruits Cx26 to lipid rafts, where it colocalizes with caveolin-1. This trafficking event appears to be unique to Cx26, since the other connexins investigated in this study do not require caveolin-1 for targeting to lipid rafts. Our results provide the first evidence that connexins interact with caveolins and partition into lipid raft domains and indicate that these interactions are connexin specific.

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The Chromatographic Efficiency of Several Commercially Available Ion Chromatography Columns

Jenke

1990

Journal of Liquid Chromatography

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The influence of mobile phase flow rate on the efficiency of six commercially available ion chromatography columns was examined. Optimum performance was achieved at flow rates between 0.3 and 0.7 mL/min. For all but one of the columns studied, the column efficiency versus reduced velocity relationship was similar to the classical Knox-type behavior. Operation at typical use conditions for all the columns examined results in performance which is only 50 to 75% as good as when the columns are operated at the flow rates optimized in terms of efficiency. Such changes in the column efficiency are directly related to the column's ability to effectively resolve sample components.

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Application of high-performance liquid chromatographic methods in the monitoring of enzyme-targeted chemotherapy

Cited by 3 publications

References 11 publications

High-performance liquid chromatographic determination and standardization of agmatine

High-performance liquid chromatographic determination and standardization of agmatine

Connexin Family Members Target to Lipid Raft Domains and Interact with Caveolin-1

The Chromatographic Efficiency of Several Commercially Available Ion Chromatography Columns

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