Abstract
The transglycosylation activity of endo-β-N-acetylglucosaminidase from Arthrobacter protophormiae (endo-A) can be enhanced dramatically by inclusion of organic solvent in the reaction mixture (see accompanying article; Fan, J.-Q., Takegawa, K., Iwahara, S., Kondo, A., Kato, I., Abeygunawardana, C., and Lee, Y. C. (1995) J. Biol. Chem. 270, 17723-17729). This finding was extended to synthesis of important intermediates for preparation of neoglycoconjugates. When 0.2 M GlcNAc-O-(CH 2 ) 6 NH 2 , GlcNAc-O-CH 2 CH=CH 2 , GlcNAc-O-(CH 2 ) 3 -CH=CH 2 , GlcNAc-O-(CH 2 ) 3 NHCOCH=CH 2 , GlcNAc-S-CH 2 CN, GlcNAc-S-(CH2) 3 CH 3 , or GlcNAc-S-CH 2 -CONHCH 2 CH(OMe) 2 were used as acceptors in 30% acetone-containing media, the transglycosylation was accomplished with about 80% yield. The transglycosylation yields to benzyl β-GlcNAc (67%), 4-methyl-umbelliferyl β-GlcNAc (66%), p-nitrophenyl β- GlcNAc (33%), and (GlcNAc-β-S-CH 2 CH 2 CH 2 ) 2 (43%) were lower, because their poor solubilities allowed only 0.05 M or lower concentrations in the reaction mixture. A micro-mole-scale synthesis of Man 9 GlcNAc 2 -O-(CH 2 ) 3 - NHCOCH=CH 2 (Man 9 GlcNAc 2 -NAP) was accomplished with 90% yield, and the structure of the transglycosylation product was confirmed by 1 H NMR. Man 9 GlcNAc 2 -NAP was co-polymerized with acrylamide. The ratio of sugar side chain to acrylamide in this glycopolymer was 1:44 and the molecular weight of glycopolymer was estimated to be between 1,500,000 and 2,000,000 by high performance gel filtration chromatography. The glyco-polymer was shown to be a much more efficient inhibitor of binding by recombinant rat mannose binding protein-carbohydrate recognition domains (MBP-CRD) from serum (I 50 = 3.5 μM Man 9 GlcNAc 2 -sugar chain) and liver (I 50 = 74.5 nM) than soybean agglutinin.
| Original language | English |
|---|---|
| Pages (from-to) | 17730-17735 |
| Number of pages | 6 |
| Journal | Journal of Biological Chemistry |
| Volume | 270 |
| Issue number | 30 |
| DOIs | |
| Publication status | Published - Jan 1 1995 |
| Externally published | Yes |
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All Science Journal Classification (ASJC) codes
- Biochemistry
- Molecular Biology
- Cell Biology
Cite this
Synthesis of neoglycoconjugates by transglycosylation with Arthrobacter protophormiae endo-β-N-acetylglucosaminidase. Demonstration of a macro- cluster effect for mannose-binding proteins. / Fan, J. Q.; Quesenberry, M. S.; Takegawa, Kaoru; Iwahara, S.; Kondo, A.; Kato, I.; Lee, Y. C.
In: Journal of Biological Chemistry, Vol. 270, No. 30, 01.01.1995, p. 17730-17735.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Synthesis of neoglycoconjugates by transglycosylation with Arthrobacter protophormiae endo-β-N-acetylglucosaminidase. Demonstration of a macro- cluster effect for mannose-binding proteins
AU - Fan, J. Q.
AU - Quesenberry, M. S.
AU - Takegawa, Kaoru
AU - Iwahara, S.
AU - Kondo, A.
AU - Kato, I.
AU - Lee, Y. C.
PY - 1995/1/1
Y1 - 1995/1/1
N2 - The transglycosylation activity of endo-β-N-acetylglucosaminidase from Arthrobacter protophormiae (endo-A) can be enhanced dramatically by inclusion of organic solvent in the reaction mixture (see accompanying article; Fan, J.-Q., Takegawa, K., Iwahara, S., Kondo, A., Kato, I., Abeygunawardana, C., and Lee, Y. C. (1995) J. Biol. Chem. 270, 17723-17729). This finding was extended to synthesis of important intermediates for preparation of neoglycoconjugates. When 0.2 M GlcNAc-O-(CH 2 ) 6 NH 2 , GlcNAc-O-CH 2 CH=CH 2 , GlcNAc-O-(CH 2 ) 3 -CH=CH 2 , GlcNAc-O-(CH 2 ) 3 NHCOCH=CH 2 , GlcNAc-S-CH 2 CN, GlcNAc-S-(CH2) 3 CH 3 , or GlcNAc-S-CH 2 -CONHCH 2 CH(OMe) 2 were used as acceptors in 30% acetone-containing media, the transglycosylation was accomplished with about 80% yield. The transglycosylation yields to benzyl β-GlcNAc (67%), 4-methyl-umbelliferyl β-GlcNAc (66%), p-nitrophenyl β- GlcNAc (33%), and (GlcNAc-β-S-CH 2 CH 2 CH 2 ) 2 (43%) were lower, because their poor solubilities allowed only 0.05 M or lower concentrations in the reaction mixture. A micro-mole-scale synthesis of Man 9 GlcNAc 2 -O-(CH 2 ) 3 - NHCOCH=CH 2 (Man 9 GlcNAc 2 -NAP) was accomplished with 90% yield, and the structure of the transglycosylation product was confirmed by 1 H NMR. Man 9 GlcNAc 2 -NAP was co-polymerized with acrylamide. The ratio of sugar side chain to acrylamide in this glycopolymer was 1:44 and the molecular weight of glycopolymer was estimated to be between 1,500,000 and 2,000,000 by high performance gel filtration chromatography. The glyco-polymer was shown to be a much more efficient inhibitor of binding by recombinant rat mannose binding protein-carbohydrate recognition domains (MBP-CRD) from serum (I 50 = 3.5 μM Man 9 GlcNAc 2 -sugar chain) and liver (I 50 = 74.5 nM) than soybean agglutinin.
AB - The transglycosylation activity of endo-β-N-acetylglucosaminidase from Arthrobacter protophormiae (endo-A) can be enhanced dramatically by inclusion of organic solvent in the reaction mixture (see accompanying article; Fan, J.-Q., Takegawa, K., Iwahara, S., Kondo, A., Kato, I., Abeygunawardana, C., and Lee, Y. C. (1995) J. Biol. Chem. 270, 17723-17729). This finding was extended to synthesis of important intermediates for preparation of neoglycoconjugates. When 0.2 M GlcNAc-O-(CH 2 ) 6 NH 2 , GlcNAc-O-CH 2 CH=CH 2 , GlcNAc-O-(CH 2 ) 3 -CH=CH 2 , GlcNAc-O-(CH 2 ) 3 NHCOCH=CH 2 , GlcNAc-S-CH 2 CN, GlcNAc-S-(CH2) 3 CH 3 , or GlcNAc-S-CH 2 -CONHCH 2 CH(OMe) 2 were used as acceptors in 30% acetone-containing media, the transglycosylation was accomplished with about 80% yield. The transglycosylation yields to benzyl β-GlcNAc (67%), 4-methyl-umbelliferyl β-GlcNAc (66%), p-nitrophenyl β- GlcNAc (33%), and (GlcNAc-β-S-CH 2 CH 2 CH 2 ) 2 (43%) were lower, because their poor solubilities allowed only 0.05 M or lower concentrations in the reaction mixture. A micro-mole-scale synthesis of Man 9 GlcNAc 2 -O-(CH 2 ) 3 - NHCOCH=CH 2 (Man 9 GlcNAc 2 -NAP) was accomplished with 90% yield, and the structure of the transglycosylation product was confirmed by 1 H NMR. Man 9 GlcNAc 2 -NAP was co-polymerized with acrylamide. The ratio of sugar side chain to acrylamide in this glycopolymer was 1:44 and the molecular weight of glycopolymer was estimated to be between 1,500,000 and 2,000,000 by high performance gel filtration chromatography. The glyco-polymer was shown to be a much more efficient inhibitor of binding by recombinant rat mannose binding protein-carbohydrate recognition domains (MBP-CRD) from serum (I 50 = 3.5 μM Man 9 GlcNAc 2 -sugar chain) and liver (I 50 = 74.5 nM) than soybean agglutinin.
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UR - http://www.scopus.com/inward/citedby.url?scp=0029023104&partnerID=8YFLogxK
U2 - 10.1074/jbc.270.30.17730
DO - 10.1074/jbc.270.30.17730
M3 - Article
C2 - 7629072
AN - SCOPUS:0029023104
VL - 270
SP - 17730
EP - 17735
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
SN - 0021-9258
IS - 30
ER -