Synthesis and characterization of a 25-residue rubredoxin(II)-like metalloprotein and its valine-leucine mutant.
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.
Christensen
H E
HE
Chemistry Department A, Technical University of Denmark, Lyngby.
Hammerstad-Pedersen
J M
JM
Holm
A
A
Roepstorff
P
P
Ulstrup
J
J
Vorm
O
O
Ostergård
S
S
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
D012416:Rubredoxins; D007930:Leucine; D014633:Valine