Abstract
We have established a facile and efficient protocol for the preparative-scale synthesis of various compound libraries related to lactosaminoglycans: cell surface oligosaccharides composed of N-acetyllactosamine as a repeating disaccharide unit, based on chemical and enzymatic approaches. Substrate specificity and feasibility of a bacterial glycosyltransferase, Neisseria meningitidis β1,3-N- acetylglucosaminyltransferase (LgtA), were investigated in order to synthesize various key intermediates suited for the construction of mammalian O-glycopeptides and glycosphingolipids containing poly-N-acetyllactosamine structures. Recombinant LgtA exhibited the highest glycosyltransferase activity with strongly basic conditions (pH = 10, glycine-NaOH buffer) and a broad range of optimal temperatures from 20 to 30 °C. Interestingly, it was found that LgtA discriminates L-serine and L-threonine and functions both as a core-1 β1,3-N-acetylglucosaminyltransferase and core-2 β1,3-N- acetylglucosaminyltransferase toward Fmoc-Ser derivatives, while LgtA showed only core-2 β1,3-N-acetylglucosaminyltransferase activity in the presence of Fmoc-Thr derivatives. Combined use of LgtA with human β1,4- galactosyltransferase allowed for controlled sugar extension reactions from synthetic sugar amino acids and gave synthetic lactosaminoglycans, such as a decasaccharide derivative, Galβ(1 → 4)GlcNAcβ(1 → 3)Galβ(1 → 4)GlcNAcβ(1 → 3)Galβ(1 → 4)GlcNAcβ(1 → 3)Galβ(1 → 4)GlcNAcβ(1 → 6)[Galβ(1 → 3)]GalNAcα1 → Fmoc-Ser-OH (6), and a dodecasaccharide derivative, Galβ(1 → 4)GlcNAcβ(1 → 3)Galβ(1 → 4)GlcNAcβ(1 → 3)Galβ(1 → 4)GlcNAcβ(1 → 6)[Galβ(1 → 4)GlcNAcβ(1 → 3)Galβ(1 → 4)GlcNAcβ(1 → 3)Galβ(1 → 3)]GalNAcα1 → Fmoc-Ser-OH (9). A partially protected pentasaccharide intermediate, GlcNAcβ(1 → 3)Galβ(1 → 4)GlcNAcβ(1 → 6)[Galβ(1 → 3)]GalNAcα1 → Fmoc-Thr-OH (11), was applied for the microwave-assisted solid-phase synthesis of a MUC1-related glycopeptide 19 (MW = 2610.1). The findings suggest that this sugar extension strategy can be employed for the modification of lactosyl ceramide mimetic polymers to afford convenient precursors for the synthesis of various glycosphingolipids.
| Original language | English |
|---|---|
| Pages (from-to) | 9609-9621 |
| Number of pages | 13 |
| Journal | Journal of Organic Chemistry |
| Volume | 71 |
| Issue number | 26 |
| DOIs | |
| Publication status | Published - Dec 22 2006 |
| Externally published | Yes |
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All Science Journal Classification (ASJC) codes
- Organic Chemistry
Cite this
Construction and structural characterization of versatile lactosaminoglycan-related compound library for the synthesis of complex glycopeptides and glycosphingolipids. / Naruchi, Kentarou; Hamamoto, Tomoki; Kurogochi, Masaki; Hinou, Hiroshi; Shimizu, Hiroki; Matsushita, Takahiko; Fujitani, Naoki; Kondo, Hirosato; Nishimura, Shin Ichiro.
In: Journal of Organic Chemistry, Vol. 71, No. 26, 22.12.2006, p. 9609-9621.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Construction and structural characterization of versatile lactosaminoglycan-related compound library for the synthesis of complex glycopeptides and glycosphingolipids
AU - Naruchi, Kentarou
AU - Hamamoto, Tomoki
AU - Kurogochi, Masaki
AU - Hinou, Hiroshi
AU - Shimizu, Hiroki
AU - Matsushita, Takahiko
AU - Fujitani, Naoki
AU - Kondo, Hirosato
AU - Nishimura, Shin Ichiro
PY - 2006/12/22
Y1 - 2006/12/22
N2 - We have established a facile and efficient protocol for the preparative-scale synthesis of various compound libraries related to lactosaminoglycans: cell surface oligosaccharides composed of N-acetyllactosamine as a repeating disaccharide unit, based on chemical and enzymatic approaches. Substrate specificity and feasibility of a bacterial glycosyltransferase, Neisseria meningitidis β1,3-N- acetylglucosaminyltransferase (LgtA), were investigated in order to synthesize various key intermediates suited for the construction of mammalian O-glycopeptides and glycosphingolipids containing poly-N-acetyllactosamine structures. Recombinant LgtA exhibited the highest glycosyltransferase activity with strongly basic conditions (pH = 10, glycine-NaOH buffer) and a broad range of optimal temperatures from 20 to 30 °C. Interestingly, it was found that LgtA discriminates L-serine and L-threonine and functions both as a core-1 β1,3-N-acetylglucosaminyltransferase and core-2 β1,3-N- acetylglucosaminyltransferase toward Fmoc-Ser derivatives, while LgtA showed only core-2 β1,3-N-acetylglucosaminyltransferase activity in the presence of Fmoc-Thr derivatives. Combined use of LgtA with human β1,4- galactosyltransferase allowed for controlled sugar extension reactions from synthetic sugar amino acids and gave synthetic lactosaminoglycans, such as a decasaccharide derivative, Galβ(1 → 4)GlcNAcβ(1 → 3)Galβ(1 → 4)GlcNAcβ(1 → 3)Galβ(1 → 4)GlcNAcβ(1 → 3)Galβ(1 → 4)GlcNAcβ(1 → 6)[Galβ(1 → 3)]GalNAcα1 → Fmoc-Ser-OH (6), and a dodecasaccharide derivative, Galβ(1 → 4)GlcNAcβ(1 → 3)Galβ(1 → 4)GlcNAcβ(1 → 3)Galβ(1 → 4)GlcNAcβ(1 → 6)[Galβ(1 → 4)GlcNAcβ(1 → 3)Galβ(1 → 4)GlcNAcβ(1 → 3)Galβ(1 → 3)]GalNAcα1 → Fmoc-Ser-OH (9). A partially protected pentasaccharide intermediate, GlcNAcβ(1 → 3)Galβ(1 → 4)GlcNAcβ(1 → 6)[Galβ(1 → 3)]GalNAcα1 → Fmoc-Thr-OH (11), was applied for the microwave-assisted solid-phase synthesis of a MUC1-related glycopeptide 19 (MW = 2610.1). The findings suggest that this sugar extension strategy can be employed for the modification of lactosyl ceramide mimetic polymers to afford convenient precursors for the synthesis of various glycosphingolipids.
AB - We have established a facile and efficient protocol for the preparative-scale synthesis of various compound libraries related to lactosaminoglycans: cell surface oligosaccharides composed of N-acetyllactosamine as a repeating disaccharide unit, based on chemical and enzymatic approaches. Substrate specificity and feasibility of a bacterial glycosyltransferase, Neisseria meningitidis β1,3-N- acetylglucosaminyltransferase (LgtA), were investigated in order to synthesize various key intermediates suited for the construction of mammalian O-glycopeptides and glycosphingolipids containing poly-N-acetyllactosamine structures. Recombinant LgtA exhibited the highest glycosyltransferase activity with strongly basic conditions (pH = 10, glycine-NaOH buffer) and a broad range of optimal temperatures from 20 to 30 °C. Interestingly, it was found that LgtA discriminates L-serine and L-threonine and functions both as a core-1 β1,3-N-acetylglucosaminyltransferase and core-2 β1,3-N- acetylglucosaminyltransferase toward Fmoc-Ser derivatives, while LgtA showed only core-2 β1,3-N-acetylglucosaminyltransferase activity in the presence of Fmoc-Thr derivatives. Combined use of LgtA with human β1,4- galactosyltransferase allowed for controlled sugar extension reactions from synthetic sugar amino acids and gave synthetic lactosaminoglycans, such as a decasaccharide derivative, Galβ(1 → 4)GlcNAcβ(1 → 3)Galβ(1 → 4)GlcNAcβ(1 → 3)Galβ(1 → 4)GlcNAcβ(1 → 3)Galβ(1 → 4)GlcNAcβ(1 → 6)[Galβ(1 → 3)]GalNAcα1 → Fmoc-Ser-OH (6), and a dodecasaccharide derivative, Galβ(1 → 4)GlcNAcβ(1 → 3)Galβ(1 → 4)GlcNAcβ(1 → 3)Galβ(1 → 4)GlcNAcβ(1 → 6)[Galβ(1 → 4)GlcNAcβ(1 → 3)Galβ(1 → 4)GlcNAcβ(1 → 3)Galβ(1 → 3)]GalNAcα1 → Fmoc-Ser-OH (9). A partially protected pentasaccharide intermediate, GlcNAcβ(1 → 3)Galβ(1 → 4)GlcNAcβ(1 → 6)[Galβ(1 → 3)]GalNAcα1 → Fmoc-Thr-OH (11), was applied for the microwave-assisted solid-phase synthesis of a MUC1-related glycopeptide 19 (MW = 2610.1). The findings suggest that this sugar extension strategy can be employed for the modification of lactosyl ceramide mimetic polymers to afford convenient precursors for the synthesis of various glycosphingolipids.
UR - http://www.scopus.com/inward/record.url?scp=33845956872&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=33845956872&partnerID=8YFLogxK
U2 - 10.1021/jo0617161
DO - 10.1021/jo0617161
M3 - Article
C2 - 17168577
AN - SCOPUS:33845956872
VL - 71
SP - 9609
EP - 9621
JO - Journal of Organic Chemistry
JF - Journal of Organic Chemistry
SN - 0022-3263
IS - 26
ER -