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Synthesis and characterization of a 25‐residue rubredoxin(II)‐like metalloprotein and its valine‐leucine mutant
Author(s) -
Christensen Hans E.M.,
Hammerstad-Pedersen Jan M.,
Holm Arne,
Roepstorff Peter,
Ulstrup Jens,
Vorm Ole,
Østergård Søren
Publication year - 1992
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(92)80939-e
Subject(s) - rubredoxin , valine , metalloprotein , leucine , chemistry , mutant , residue (chemistry) , biochemistry , stereochemistry , amino acid , enzyme , gene
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 → Leu 41 chemical mutant has also been synthesized and behaves in a closely similar fashion.