title: Poly(3-hydroxybutyrate) is associated with specific proteins in the cytoplasm and membranes of Escherichia coli.
abstract: Poly(3-hydroxybutyrate) (PHB) is well-known as a high molecular weight homopolymer of R-3-hydroxybutyrate which accumulates in storage granules within the cytosols of certain bacteria. Escherichia coli does not amass these granules; however, small amounts of low molecular weight PHB (<0.02% of dry weight) have been found in the plasma membranes in complexes with calcium polyphosphate; the complexes serve as voltage-activated calcium channels. Here we report that polyphosphate-complexed PHB is only a minor fraction of the polyester in E. coli. PHB comprises 0.36 to 0. 55% of the dry weight of log-phase cells, depending on culture medium, and this amount increases by 15 to 20% when the cells are made genetically competent. The PHB is widely distributed throughout the cell, wherein it is primarily associated with proteins. The identity of protein-associated PHB was established by antibody reaction, chemical assay, and 1H NMR spectroscopy. As expected, the physical and chemical properties of protein-associated PHB were found to be considerably different from those of the bulk polymer or granule PHB, e.g. protein-PHB complexes are normally insoluble in chloroform, soluble in water and alkaline hypochlorite, and are converted to crotonic acid more slowly on heating in concentrated sulfuric acid. Our studies indicate that the majority of cellular PHB (over 80%) is located in cytoplasmic proteins, especially proteins of the ribosomal fraction. Western immunoblots, probed with polyclonal anti-PHB IgG, revealed a number of PHB-polypeptides having a wide range of molecular weights in all cell fractions. These results suggest that PHB is a fundamental constituent of cells that may have physiological functions in addition to facilitating ion transmembrane transport or serving as a carbon reserve.
authors.affiliation: Department of Microbiology, Michigan State University, East Lansing, Michigan 48824, USA.
authors.forename: R
authors.initials: R
authors.lastname: Huang
authors.forename: R N
authors.initials: RN
authors.lastname: Reusch
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism*; D015153:Blotting, Western; D002462:Cell Membrane / Q000378:metabolism; D003437:Crotonates / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism*; D006885:Hydroxybutyrates / Q000302:isolation & purification / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D008970:Molecular Weight; D011091:Polyesters / Q000302:isolation & purification / Q000378:metabolism*; D013464:Sulfuric Acids / Q000378:metabolism; D000094122:Polyhydroxybutyrates
chemical_list: D001426:Bacterial Proteins; D003437:Crotonates; D006885:Hydroxybutyrates; D011091:Polyesters; D013464:Sulfuric Acids; C003182:poly-beta-hydroxybutyrate; D000094122:Polyhydroxybutyrates; C033158:sulfuric acid