Journal Issue - Volume 7 Issue 8 (August 1998)
Aromatic L-amino acid decarboxylase: Conformational change in the flexible region around arg334 is required during the transaldimination process
- Seiji Ishii, Hideyuki Hayashi, Akihiro Okamoto, Hiroyuki Kagamiyama
- Published in Wiley Interscience on Dec 31, 2008
- DOI: 10.1002/pro.5560070816 (p 1802-1810)
For further analyses, we constructed a mutant gene encoding in tandem the two peptides of AADC cleaved at the Asn327-Met328 bond inside the putative flexible region. The gene product, fragmentary AADC, was still active with L-dopa as substrate, but its kcar value was decreased 57-fold, and the Km value was increased 9-fold compared with those of the wild-type AADC. The absorption spectra of the fragmentary AADC in the presence of L-dopa methyl ester showed shift in the equilibrium of the...
Opposite behavior of two isozymes when refolding in the presence of non-ionic detergents
- Fernando Doñate, Antonio Artigues, Ana Iriarte, Marino Martinez-Carrion
- Published in Wiley Interscience on Dec 31, 2008
- DOI: 10.1002/pro.5560070817 (p 1811-1820)
GroEL has a greater affinity for the mitochondrial isozyme (mAAT) of aspartate aminotransferase than for its cytosolic counterpart (cAAT) (Mattingly JR Jr, Iriarte A, Martinez-Carrion M, 1995,J Biol Chem 270:1138-1148), two proteins that share a high degree of sequence similarity and an almost identical spatial structure. The effect of detergents on the refolding of these large, dimeric isozymes parallels this difference in behavior. The presence of non-ionic detergents such as Triton X-100 or...
Turn scanning by site-directed mutagenesis: Application to the protein folding problem using the intestinal fatty acid binding protein
- Keehyuk Kim, Carl Frieden
- Published in Wiley Interscience on Dec 31, 2008
- DOI: 10.1002/pro.5560070818 (p 1821-1828)
We have systematically mutated residues located in turns between -strands of the intestinal fatty acid binding protein (IFABP), and a glycine in a half turn, to valine and have examined the stability, refolding rate constants and ligand dissociation constants for each mutant protein. IFABP is an almost all -sheet protein exhibiting a topology comprised of two five-stranded sheets surrounding a large cavity into which the fatty acid ligand binds. A glycine residue is located in seven of the...
A superfamily of metalloenzymes unifies phosphopentomutase and cofactor-independent phosphoglycerate mutase with alkaline phosphatases and sulfatases
- Michael Y. Galperin, Eugene V. Koonin, Amos Bairoch
- Published in Wiley Interscience on Dec 31, 2008
- DOI: 10.1002/pro.5560070819 (p 1829-1835)
Sequence analysis of the probable archaeal phosphoglycerate mutase resulted in the identification of a superfamily of metalloenzymes with similar metal-binding sites and predicted conserved structural fold. This superfamily unites alkaline phosphatase, N-acetylgalactosamine-4-sulfatase, and cerebroside sulfatase, enzymes with known three-dimensional structures, with phosphopentomutase, 2,3-bisphosphoglycerate-independent phosphoglycerate mutase, phosphoglycerol transferase, phosphonate...
Crystallization of recombinant human heme oxygenase-1
- David J. Schuller, Thomas L. Poulos, Angela Wilks, Paul Ortiz De Montellano
- Published in Wiley Interscience on Dec 31, 2008
- DOI: 10.1002/pro.5560070820 (p 1836-1838)
Heme oxygenase catalyzes the NADPH, 02, and cytochrome P450 reductase dependent oxidation of heme to biliverdin and carbon monoxide. One of two primary isozymes, HO-1, is anchored to the endoplasmic reticulum membrane via a stretch of hydrophobic residues at the C-terminus. While full-length human HO-1 consists of 288 residues, a truncated version with residues 1-265 has been expressed as a soluble active enzyme in Escherichia coli. The recombinant enzyme crystallized from ammonium sulfate...
Overexpression of recombinant proteins with a C-terminal thiocarboxylate: Implications for protein semisynthesis and thiamin biosynthesis
- Cynthia Kinsland, Sean V. Taylor, Neil L. Kelleher, Fred W. Mclafferty, Tadhg P. Begley
- Published in Wiley Interscience on Dec 31, 2008
- DOI: 10.1002/pro.5560070821 (p 1839-1842)
A facile and rapid method for the production of protein C-terminal thiocarboxylates on DNA-encoded polypeptides is described. This method, which relies on the mechanism of the cleavage reaction of intein-containing fusion proteins, can produce multi-milligram quantities of protein C-terminal thiocarboxylate quickly and inexpensively. The utility of this method for protein semisynthesis and implications for studies on the biosynthesis of thiamin are discussed.
Erratum
- Published in Wiley Interscience on Dec 31, 2008
- DOI: 10.1002/pro.5560070822 (p 1843)
No Abstract.



