| RRC ID |
922
|
| Author |
Saskiawan I, Mizuno M, Inazu T, Haneda K, Harashima S, Kumagai H, Yamamoto K.
|
| Title |
Chemo-enzymatic synthesis of the glycosylated alpha-mating factor of Saccharomyces cerevisiae and analysis of its biological activity.
|
| Journal |
Arch Biochem Biophys
|
| Abstract |
The effect of glycosylation on a bioactive peptide was studied using yeast Saccharomyces cerevisiae alpha-mating factor, which is composed of 13 amino acids. In this study, we prepared glycosylated alpha-mating factor by chemo-enzymatic synthesis. At first, N-acetylglucosaminyl alpha-mating factor (Trp-His-Trp-Leu-Gln(GlcNAc)-Leu-Lys-Pro-Gly-Gln-Pro-Met-Tyr) was chemically synthesized by the solid-phase method. Then, using the transglycosylation activity of Mucor hiemalis endo-beta-N-acetylglucosaminidase, we synthesized glycosylated alpha-mating factor with a glutamine-linked sialo complex type oligosaccharide. The biological activity of alpha-mating factor derivatives was examined by means of a growth arrest assay using secreted-protease-defective a cells of S. cerevisiae. The results showed that the bioactivity of glycosylated alpha-mating factor was lower than that of native alpha-mating factor. However, when sialic acid was removed from the complex type sugar chain of glycosylated alpha-mating factor, its bioactivity was recovered. Glycosylated alpha-mating factor exhibited higher resistance against proteolysis than native alpha-mating factor. It was found that the bioactivity of N-acetylglucosaminyl alpha-mating factor was higher than that of alpha-mating factor. Circular dichroism studies indicated that a slight change in the structure of alpha-mating factor may influence its activity.
|
| Volume |
406(1)
|
| Pages |
127-34
|
| Published |
2002-10-1
|
| DOI |
10.1016/s0003-9861(02)00416-2
|
| PII |
S0003986102004162
|
| PMID |
12234498
|
| MeSH |
Acetylglucosamine / metabolism*
Agglutinins / biosynthesis
Amino Acid Sequence
Circular Dichroism
Glycoside Hydrolases / metabolism
Glycosylation*
Mating Factor
Mucor / metabolism
Peptides / chemical synthesis*
Peptides / chemistry
Peptides / metabolism*
Protein Conformation
Saccharomyces cerevisiae / genetics
Saccharomyces cerevisiae / growth & development
Saccharomyces cerevisiae / metabolism*
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
|
| IF |
3.391
|
| Times Cited |
20
|
|
WOS Category
|
BIOPHYSICS
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| Resource |
| Yeast |
BY1658(SH1658) |
