Interaction of protein-bound polysaccharide (PSK) with smooth muscle myosin regulatory light chain.
The interaction of a protein-bound polysaccharide (PSK) isolated from Basidiomycetes with smooth muscle myosin components was evaluated by limited digestion, urea/glycerol gel electrophoresis, affinity chromatography and overlay assay using a peptide array. PSK was bound to the regulatory light chain (RLC) of myosin, but not to the essential light chain. The binding to PSK was definitely observed for unphosphorylated RLC, compared to phosphorylated one. From the amino acid sequence of the RLC, 490 peptides were synthesized on a cellulose membrane. Overlay assays showed that the PSK-binding on the molecule of RLC were localized in the N- and C-terminal basic regions and these sites were conserved in RLC from the human smooth muscle and nonmuscle cells.
Fujii
Toshihiro
T
Department of Kansei Engineering, Faculty of Textile Science and Technology, Shinshu University, Ueda, Nagano 386-8567, Japan. fujiit1@giptc.shinshu-u.ac.jp
Kunimatsu
Mitoshi
M
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D002645:Chickens; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D005895:Gizzard, Avian / Q000737:chemistry / Q000201:enzymology; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D009130:Muscle, Smooth / Q000737:chemistry* / Q000201:enzymology; D009219:Myosin-Light-Chain Kinase / Q000737:chemistry* / Q000302:isolation & purification; D009218:Myosins / Q000737:chemistry* / Q000302:isolation & purification; D009842:Oligopeptides / Q000737:chemistry; D010766:Phosphorylation; D040081:Protein Array Analysis; D011487:Protein Conformation; D011509:Proteoglycans / Q000737:chemistry*
D009842:Oligopeptides; D011509:Proteoglycans; C010770:polysaccharide-K; D009219:Myosin-Light-Chain Kinase; D009218:Myosins