TY - JOUR
T1 - UV Raman monitoring of histidine protonation and H-2H exchange in plastocyanin
AU - Wu, Qiang
AU - Li, Fangbiao
AU - Wang, Weixun
AU - Hecht, Michael H.
AU - Spiro, Thomas G.
N1 - Funding Information:
This work was supported by NIH Grant GM 33576 from the Institute of General Medical Sciences.
PY - 2002
Y1 - 2002
N2 - UV resonance Raman bands of Cu-bound and protonated histidine residues have been detected in 2H2O solutions of poplar plastocyanin. For the Cu(II) protein, slow NH-2H exchange of the His37 ligand was monitored via the growth of bands at 1389 and 1344 cm-1 when Pcy was exchanged into 2H2O, or via their diminution when the protein was exchanged back into H2O; the rate constant is 7×10-4/s at pH (p2H) 7.4 at room temperature. The slow exchange is attributed to imidazole H-bonding to a backbone carbonyl. Nearby bands at 1397 and 1354 cm-1, appear and disappear within the mixing time, and are assigned to the solvent-exposed His87 ligand. The ∼10 cm-1 differences between His37 and His87 are attributed to the effect of H-bonding on the imidazole ring modes. The UVRR spectra of the Cu(I) protein in 2H2O reveal a 1408 cm-1 band, characteristic of NH-2H-exchanged histidinium, which grows in as the p2H is lowered. Its intensity follows a titration curve with pKa=4.6. This protonation is assigned to the His87 residue, whose bond to the Cu(I) is known from crystallography to be broken at low pH. As the 1408 cm-1 band grows, a band at 1345 cm-1 diminishes, while another, at 1337 cm-1 stays constant. These are assigned to modes of bound His87 and His37, respectively, shifted down 7-9 cm-1 from their Cu(II) positions.
AB - UV resonance Raman bands of Cu-bound and protonated histidine residues have been detected in 2H2O solutions of poplar plastocyanin. For the Cu(II) protein, slow NH-2H exchange of the His37 ligand was monitored via the growth of bands at 1389 and 1344 cm-1 when Pcy was exchanged into 2H2O, or via their diminution when the protein was exchanged back into H2O; the rate constant is 7×10-4/s at pH (p2H) 7.4 at room temperature. The slow exchange is attributed to imidazole H-bonding to a backbone carbonyl. Nearby bands at 1397 and 1354 cm-1, appear and disappear within the mixing time, and are assigned to the solvent-exposed His87 ligand. The ∼10 cm-1 differences between His37 and His87 are attributed to the effect of H-bonding on the imidazole ring modes. The UVRR spectra of the Cu(I) protein in 2H2O reveal a 1408 cm-1 band, characteristic of NH-2H-exchanged histidinium, which grows in as the p2H is lowered. Its intensity follows a titration curve with pKa=4.6. This protonation is assigned to the His87 residue, whose bond to the Cu(I) is known from crystallography to be broken at low pH. As the 1408 cm-1 band grows, a band at 1345 cm-1 diminishes, while another, at 1337 cm-1 stays constant. These are assigned to modes of bound His87 and His37, respectively, shifted down 7-9 cm-1 from their Cu(II) positions.
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U2 - 10.1016/S0162-0134(01)00354-3
DO - 10.1016/S0162-0134(01)00354-3
M3 - Article
C2 - 11897354
AN - SCOPUS:0036186024
SN - 0162-0134
VL - 88
SP - 381
EP - 387
JO - Journal of Inorganic Biochemistry
JF - Journal of Inorganic Biochemistry
IS - 3-4
ER -