Title: | Enhancement of bioactivity of Saccharomyces cerevisiae alpha-mating factor by attachment of sugar moiety to glutamine residue |
Author(s): | Saskiawan I; Mizuno M; Inazu T; Haneda K; Kumagai H; Yamamoto K; |
Address: | "Graduate School of Biostudies, Kyoto University, Kitashirakawa, Sakyo-ku, Kyoto 606-8502, Japan" |
DOI: | 10.1016/j.jbiotec.2004.07.005 |
ISSN/ISBN: | 0168-1656 (Print) 0168-1656 (Linking) |
Abstract: | "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" |
Keywords: | Acetylglucosamine/chemistry Acetylglucosaminidase/metabolism Biochemistry/methods Carbohydrate Sequence Circular Dichroism Glucose/chemistry Glutamine/*chemistry Molecular Sequence Data Protein Engineering/*methods Protein Precursors/*chemistry/metabolism; |
Notes: | "MedlineSaskiawan, Iwan Mizuno, Mamoru Inazu, Toshiyuki Haneda, Katsuji Kumagai, Hidehiko Yamamoto, Kenji eng Research Support, Non-U.S. Gov't Netherlands 2004/11/04 J Biotechnol. 2004 Nov 9; 114(3):299-306. doi: 10.1016/j.jbiotec.2004.07.005" |