Alkaline transition of phytocyanins: a comparison of stellacyanin and umecyanin.
The effect of pH on Cu(I) and Cu(II) forms of the isolated soluble domain of the stellacyanin from Rhus vernicifera (SCu), the Japanese lacquer tree, has been studied by electronic and NMR spectroscopy and using direct electrochemical measurements. A pK(a) value of 10.1-10.4 is observed for the alkaline transition in this oxidized phytocyanin and results in a slightly altered active-site structure, as indicated by changes in the visible and paramagnetic (1)H NMR spectra. Electrochemical studies show that the pK(a) value for this transition in SCu(I) (reduced SCu) is 11.0. These results are compared with those recently obtained for other phytocyanins, and in particular umecyanin. In all cases, the alkaline transition is caused by the deprotonation of the surface lysine residue adjacent to the axial ligand. This lysine residue is completely conserved in known phytocyanin sequences. Also highlighted in these studies are the remarkable active-site similarities between stellacyanin and umecyanin.
Dennison
Christopher
C
School of Natural Sciences, University of Newcastle upon Tyne, Newcastle upon Tyne NE1 7RU, UK. christopher.dennison@ncl.ac.uk
Harrison
Mark D
MD
Lawler
Anne T
AT
D000595:Amino Acid Sequence; D003300:Copper; D006863:Hydrogen-Ion Concentration; D007564:Japan; D009682:Magnetic Resonance Spectroscopy; D008667:Metalloproteins / Q000737:chemistry* / Q000302:isolation & purification; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000737:chemistry* / Q000302:isolation & purification; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D029045:Rhus / Q000737:chemistry; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013053:Spectrophotometry
D008667:Metalloproteins; D010940:Plant Proteins; C002054:umecyanin; C003343:stellacyanin protein, plant; D003300:Copper