title: Synthesis and characterization of a 25-residue rubredoxin(II)-like metalloprotein and its valine-leucine mutant.
abstract: An iron-sulfur metalloprotein containing the 5-12 and 35-50 residues of Desulfovibrio 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 of 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.
authors.affiliation: Chemistry Department A, Technical University of Denmark, Lyngby.
authors.forename: H E
authors.initials: HE
authors.lastname: Christensen
authors.forename: J M
authors.initials: JM
authors.lastname: Hammerstad-Pedersen
authors.forename: A
authors.initials: A
authors.lastname: Holm
authors.forename: P
authors.initials: P
authors.lastname: Roepstorff
authors.forename: J
authors.initials: J
authors.lastname: Ulstrup
authors.forename: O
authors.initials: O
authors.lastname: Vorm
authors.forename: S
authors.initials: S
authors.lastname: Ostergård
mesh_terms: D000595:Amino Acid Sequence; D007930:Leucine / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation; D012416:Rubredoxins / Q000096:biosynthesis* / Q000737:chemistry / Q000378:metabolism; D013057:Spectrum Analysis; D014633:Valine / Q000235:genetics
chemical_list: D012416:Rubredoxins; D007930:Leucine; D014633:Valine