Synthesis and characterization of a 25‐residue rubredoxin(II)‐like metalloprotein and its valine‐leucine mutant

FEBS Letters

Hammerstad-Pedersen, Jan M.¹  Christensen, Hans E.M.¹  Vorm, Ole³  Holm, Arne²  Roepstorff, Peter³  Ulstrup, Jens¹  Østergård, Søren² 

[1] Chemistry Department A, Building 207, The Technical University of Denmark, 2800 Lyngby, Denmark;Department of Chemistry, Centre for Medical Biotechnology, The Royal Veterinary and Agricultural University, 1871 Copenhagen, Denmark;Department of Molecular Biology, Odense University, 5230 Odense, Denmark

关键词: Metalloprotein;    Rubredoxin;    Synthesis;    Characterization;    Chemical mutant;   

DOI  :  10.1016/0014-5793(92)80939-E

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

An iron-sulfur metalloprotein containing the 5–12 and 35–50 residues of Desulfovlbrio gigas rubredoxin has been synthesized by Fmoc solid phase peptide synthesis and subsequent peptide folding. A Gly links the two residue chains between Val-5 and Glu-50. Sybyl Tripos structure optimization indicates only minor structural changes of the folded synthetic protein compared to the similar residue positions in the native protein. The UV-VIS spectrum of the reduced synthetic protein is very similar to that or native D. gigas rubredoxin and the molecular mass determined by laser mass spectrometry has the expected value (± 2D). No metal is transferred to the gas phase by the laser beam merely by mixing the peptide and iron(II), substantiating that the folding procedure is a necessary pre-requisite for protein formation. The Val → Leu⁴¹ chemical mutant has also been synthesized and behaves in a closely similar fashion.

【 授权许可】

Unknown