Regulation of fungal cell wall growth: a guanine nucleotide-binding, proteinaceous component required for activity of (1----3)-beta-D-glucan synthase.
By treatment with detergent and NaCl, particulate (1----3)-beta-D-glucan synthase (EC 2.4.1.34) from Hansenula anomala or Neurospora crassa was dissociated into a "soluble fraction" and a "membrane fraction." Each fraction alone was almost inactive, but enzymatic activity could be reconstituted by mixing the two fractions and adding GTP or one of its analogs. Based on their lability to heat and to incubation with trypsin, the activity in both fractions is proteinaceous. The active component in the soluble fraction appears to bind guanosine 5'-[gamma-thio]triphosphate (GTP[gamma S]), since it was specifically protected by this nucleotide against heat inactivation and against inactivation in the presence of EDTA. Furthermore, precipitation of the soluble component with ammonium sulfate in the presence of GTP[gamma S] gave rise to a fraction that was highly active in the absence of added nucleotide, indicating either tight binding or covalent interaction between GTP[gamma S] and the soluble component. The membrane fraction probably contains the catalytic moiety, because it was partially protected against heat inactivation by the substrate, UDP-glucose. Soluble fractions that stimulated membrane fractions from H. anomala and N. crassa were obtained from several other fungi, including Saccharomyces cerevisiae. We propose that the soluble fraction contains a GTP-binding protein that modulates the biosynthesis of (1----3)-beta-D-glucan of fungal cell walls and probably has a major role in the regulation of cell wall morphogenesis.
D002462:Cell Membrane / Q000201:enzymology; D002473:Cell Wall / Q000502:physiology; D005658:Fungi / Q000201:enzymology*; D019204:GTP-Binding Proteins / Q000378:metabolism*; D005964:Glucosyltransferases / Q000378:metabolism*; D007700:Kinetics; D008565:Membrane Proteins; D009491:Neurospora / Q000201:enzymology*; D009492:Neurospora crassa / Q000201:enzymology* / Q000502:physiology; D010843:Pichia / Q000201:enzymology* / Q000502:physiology; D004718:Saccharomycetales / Q000201:enzymology*; D029702:Schizosaccharomyces pombe Proteins; D013045:Species Specificity
D008565:Membrane Proteins; D029702:Schizosaccharomyces pombe Proteins; D005964:Glucosyltransferases; C020570:1,3-beta-glucan synthase; D019204:GTP-Binding Proteins