Enhancement of bioactivity of Saccharomyces cerevisiae alpha-mating factor by attachment of sugar moiety to glutamine residue.
We prepared yeast Saccharomyces cerevisiae alpha-mating factor, a 13-amino acid pheromone produced by haploid alpha-cells, bound with glucose or N-acetylglucosamine at the fifth glutamine residue from the N-terminal by the chemical method of peptide synthesis. It was found that the bioactivity of glucosyl alpha-mating factor was higher than that of native alpha-mating factor. However, it was slightly lower than that of N-acetylglucosaminyl alpha-mating factor. This suggested that the N-acetylamino residue might play some important role in the enhancement of the bioactivity of alpha-mating factor. However, CD spectra analysis of alpha-mating factor and its derivatives demonstrated that their structures were almost identical. On the other hand, we attached a sialo complex type oligosaccharide to N-acetylglucosamine or its glucose residue by means of the transglycosylation activity of endo-beta-N-acetylglucosaminidase from Mucor hiemalis (Endo-M). The attachment of the oligosaccharide to both alpha-mating factors reduced their activities. However, enzymatical trimming of the sialo complex type oligosaccharide recovered its activity.
Saskiawan
Iwan
I
Graduate School of Biostudies, Kyoto University, Kitashirakawa, Sakyo-ku, Kyoto 606-8502, Japan.
Mizuno
Mamoru
M
Inazu
Toshiyuki
T
Haneda
Katsuji
K
Kumagai
Hidehiko
H
Yamamoto
Kenji
K
D000117:Acetylglucosamine / Q000737:chemistry; D000118:Acetylglucosaminidase / Q000378:metabolism; D001671:Biochemistry / Q000379:methods; D002240:Carbohydrate Sequence; D002942:Circular Dichroism; D005947:Glucose / Q000737:chemistry; D005973:Glutamine / Q000737:chemistry*; D008969:Molecular Sequence Data; D015202:Protein Engineering / Q000379:methods*; D011498:Protein Precursors / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000187:drug effects; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology*; D013329:Structure-Activity Relationship
C048632:MF(ALPHA)1 protein, S cerevisiae; D011498:Protein Precursors; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; D005973:Glutamine; D000118:Acetylglucosaminidase; D005947:Glucose; D000117:Acetylglucosamine