Screening for beta-poly(L-malate) binding proteins by affinity chromatography.

Thomas Göttler; Eggehard Holler

doi:10.1016/j.bbrc.2006.01.064

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Screening for beta-poly(L-malate) binding proteins by affinity chromatography.

Thomas Göttler·Eggehard Holler

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Abstract

Poly(beta-L-malic acid) is a cell type-specific polymer of myxomycetes (true slime molds) with the physiological role to organize mobility of certain proteins over the giant multinucleated plasmodia. We have developed an affinity chromatography employing 1,6-diamino-n-hexane-Sepharose-coupled poly(malic acid) to identify such proteins in cellular extracts of Physarum polycephalum. Molecular masses were measured by SDS-PAGE and non-denaturing PAGE after silver staining and/or Western blotting. Protein complexes/subunits were detected by 2-dimensional non-denaturing PAGE/SDS-PAGE. A simplified gel shift experiment displayed binding to fragmented calf thymus DNA. Nuclei were richest in poly(malate) binding proteins followed by cytoplasm and membranes. A protein of 370 kDa dissociated into 11 subunits of 11-29 kDa, indicative of a highly complex protein. This and other proteins displayed binding to nucleic acid in gel shift experiments. Poly(malate) is considered a structural and functional equivalent of long contiguous aspartate repeats in proteins of eukaryotes.

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Animals Blotting, Western Carrier Proteins Chromatography, Affinity Chromatography, Gel Diamines Electrophoresis, Polyacrylamide Gel Malates Molecular Weight Physarum polycephalum Polymers Sepharose Journal Article

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Thomas Göttler, Eggehard Holler. 2006-01-26. Screening for beta-poly(L-malate) binding proteins by affinity chromatography.. https://doi.org/10.1016/j.bbrc.2006.01.064

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