PubMed Health⌕ Search

Biomedical subjects

C K Watanabe

Publications and source records attributed to C K Watanabe.

4 recordsLinked to original sources

Rapid mapping of protein functional epitopes by combinatorial alanine scanning.

A combinatorial alanine-scanning strategy was used to determine simultaneously the functional contributions of 19 side chains buried at the interface between human growth hormone and the extracellular domain of its receptor. A phage-displayed protein library was constructed in which the 19 side chains were preferentially allowed to vary only as the wild type or alanine. The library pool was subjected to binding selections to isolate functional clones, and DNA sequencing was used to determine the alanine/wild-type ratio at each varied position. This ratio was used to calculate the effect of each alanine substitution as a change in free energy relative to that of wild type. Only seven side chains contribute significantly to the binding interaction, and these conserved residues form a compact cluster in the human growth hormone tertiary structure. The results were in excellent agreement with free energy data previously determined by conventional alanine-scanning mutagenesis and suggest that this technology should be useful for analyzing functional epitopes in proteins.

Alanine↗

Compilation and comparison of the sequence context around the AUG startcodons in Saccharomyces cerevisiae mRNAs.

The nucleotide sequence of the translation initiation regions of 96 Saccharomyces cerevisiae mRNAs was compiled and compared. The entire 5' untranslated sequence of most mRNAs is very rich in A-residues. G-residues are underrepresented in the untranslated region. The AUG startcodon context appeared to be distinctly different from that of animal mRNAs, although an A-residue at -3 also occurs very frequently (81 percent) in yeast mRNAs. The prevailing codon 3' adjacent to the AUG is the UCU serine codon. All these features are more extreme in the highly expressed genes. Fifty percent of all highly expressed genes use the UCU serine codon as second triplet. In this group G-residues are completely absent in the 7 bases preceding the startcodon and an A-residue occurs at position -1 and -3 at a frequency of 89 percent and 100 percent, respectively. The abundance of A-residues throughout the leader suggests that unstructured mRNA is required for efficient translation initiation in yeast. The consensus sequence for the AUG context in highly expressed genes can be summarized as follows: (Sequence: see text).

Base Sequence↗

Familial centronuclear myopathy.

The clinical and histological features of two Negro brothers with a centronuclear myopathy are described. They bring to 19 the number of cases now reported with this constellation of physical signs and pathological changes in the muscles. A review of these patients suggests the existence of several different diseases causing this picture, though presumably the underlying biochemical defects are closely related. It is concluded that these myopathies are degenerative rather than due to arrest of foetal muscle maturation.

Adult↗