Nuclear magnetic resonance determination of intramolecular distances in bovine pancreatic trypsin inhibitor using nitrotyrosine chelation of lanthanides

Biochemistry
T D MarinettiB D Sykes

Abstract

Nitration of tyrosine has been investigated as a means for chemically introducing lanthanide chelating sites at known positions in proteins. The low-field portions of the 250-MHZ and 270-MHZ 1H nuclear magnetic resonance spectra of native and chemically modified bovine pancreatic trypsin inhibitor have been studied in the presence of lanthanide ions. Comparisons of spectral changes observed with native, mononitro (tryosine 10) and dinitro (tyrosines 10 and 21) derivatives enable these changes to be separately attributed to metal bound at nitrotyrosine 21, nitrotyrosine 10, or the set of five carboxyl groups. The pH dependence of Pr(III) and Eu(III) induced chemical shifts yields stability constants of 50 and 159 M-1 for the association between lanthanides and nitrotyrosines 10 and 21, respectively. Correlation times for the interactions with Gd(III) bound to specific nitrotyrosines are estimated from the induced line broadening of resonances of the nitrotyrosine ring protons. These stability constants and correlation times are used to determine the distances from the different metal binding sites to buried backbone NH protons having resolved resonances. Comparisons with distances in the x-ray crystal structure give assignments ...Continue Reading

References

Jun 17, 1975·Proceedings of the Royal Society of London. Series B, Containing Papers of a Biological Character·I D CampbellJ P Williams
Dec 31, 1973·Annals of the New York Academy of Sciences·I D CampbellA V Xavier
Mar 14, 1972·Biochimica Et Biophysica Acta·C D BarryA V Xavier
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Mar 1, 1968·European Journal of Biochemistry·B MelounF Sorm
Jan 1, 1972·Methods in Enzymology·J F Riordan, B L Vallee

Citations

Nov 1, 1983·Quarterly Reviews of Biophysics·S W Englander, N R Kallenbach
Mar 15, 1978·European Journal of Biochemistry·J H BradburyB Warren
Dec 17, 1979·European Journal of Biochemistry·T EndoT Miyazawa
Jan 1, 1981·International Journal of Peptide and Protein Research·R WalterV J Hruby

Related Concepts

Metazoa
Bos indicus
Hydrogen-Ion Concentration
Mathematics
Metals, Rare Earth
In Vivo NMR Spectroscopy
Plasma Protein Binding Capacity
Protein Conformation
Pancreatic Trypsin Secretory Inhibitor, Kazal
Trypsin Inhibitors

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