PMID: 11606211Oct 19, 2001Paper

Reaction of hen egg white lysozyme with Fischer-type metallocarbene complexes. Characterization of the conjugates and determination of the metal complex binding sites

European Journal of Biochemistry
M SalmainGérard Jaouen

Abstract

The introduction of heavy atoms into protein crystals is sometimes rendered difficult and tedious because of the poor specificity of the available reagents for particular target residues. On the other hand, transition organometallic chemistry offers an almost untouched field for this purpose. In particular, Fischer-type metallocarbene complexes of the general formula (CO)5W=C(OR1)R2 may be attractive reagents because they contain the heavy element tungsten and specifically target amino groups to form stable, covalent aminocarbene adducts. With a small protein such as hen egg white lysozyme (HEWL) with a limited number of potential binding sites, it was possible to form protein-aminocarbene conjugates that have an average of one aminocarbene moiety per protein molecule. RP-HPLC combined with matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) MS analysis of the conjugates revealed that they were mixtures of the native protein, monoaminocarbenes and diaminocarbenes. Tryptic proteolysis experiments performed on the protein conjugates combined with MALDI-TOF-MS analysis of the aminocarbenic peptides allowed us to determine that lysines 13, 33, 97 and 116 were involved in the reaction of HEWL with (CO)5W=C(OMe)Me.

References

Jun 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·D SuckauM Przybylski
May 1, 1989·Analytical Biochemistry·J Trudel, A Asselin
Sep 15, 1973·Journal of Molecular Biology·A Shrake, J A Rupley
May 18, 1981·Carbohydrate Research·H Yamada, T Imoto

❮ Previous
Next ❯

Citations

Mar 23, 2010·Analytical and Bioanalytical Chemistry·Susanne BomkeUwe Karst
Dec 15, 2006·Chembiochem : a European Journal of Chemical Biology·Pierre HaquetteGérard Jaouen
Aug 2, 2011·Metallomics : Integrated Biometal Science·Sarah L BinkleyChristopher J Ziegler
May 8, 2010·Chemical Communications : Chem Comm·Sarah L BinkleyRoger S Rowlett

❮ Previous
Next ❯

Related Concepts

Related Feeds

Bacterial Cell Wall Structure

Bacterial cell walls are made of peptidoglycan (also called murein), which is made from polysaccharide chains cross-linked by unusual peptides containing D-amino acids. Here is the latest research on bacterial cell wall structures.

Bacterial Cell Wall Structure (ASM)

Bacterial cell walls are made of peptidoglycan (also called murein), which is made from polysaccharide chains cross-linked by unusual peptides containing D-amino acids. Here is the latest research on bacterial cell wall structures.