Structures of the N-linked sugar chains in PAS-7 glycoprotein sharing the same protein core with PAS-6 glycoprotein from the bovine milk fat globule membrane

Jin Seok Seok, Michiko Shimoda, Norihiro Azuma, Choemon Kanno

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Glycoproteins PAS-6 (50 kDa) and -7 (47 kDa) from the bovine milk fat globule membrane share a common protein core but differ in their carbohydrate moiety. We here analyzed and proposed the structures of the N-linked sugar chains of PAS-7. The N-linked sugar chains were liberated from PAS-7 by hydrazinolysis and, after modifying the reducing ends with 2-aminopyridine (PA), were separated into one neutral (7N, 55%) and two acidic (7M, mono-, 43%; 7D, di-, 2%) sugar chain groups. The latter were converted into neutral groups (7MN and 7DN) by sialidase digestion. 7N was finally separated into 5 chains (7N1A, 7N1B-1, 7N1B-2, 7N2A, and 7N2B), and 7MN and 7DN were separated into 3 (7MN1, 7MN2, and 7MN3) and 2 (7DN1 and 7DN2) chains, respectively. The structure of each of these PA-neutral sugar chains was determined by sugar analysis, sequential exoglycosidase digestion, partial acetolysis, and 1H-NMR spectroscopy. The results show that the 10 sugar chains were of the biantennary complex type with and without fucose. The structure of 7N2A, one of the major sugar chains, was proposed as; Galβ1-4GlcNAcβ1-2Manα1↘ 63 Manβ1-4GlcNAcβ1-4GlcNAc-PA Galβ1-4GlcNAcβ1-2Manα1↗ A structural comparison between PAS-6 and -7 indicated that, although they shared the same protein core, their sugar moiety was markedly different, involving the existence of a different pathway during the post-transcriptional modification.

Original languageEnglish (US)
Pages (from-to)901-912
Number of pages12
JournalBioscience, Biotechnology and Biochemistry
Volume65
Issue number4
DOIs
StatePublished - Apr 1 2001
Externally publishedYes

Fingerprint

Sugars
Digestion
Glycoproteins
Membranes
Fucose
Glycoside Hydrolases
Neuraminidase
Proteins
Magnetic Resonance Spectroscopy
Carbohydrates
milk fat globule
Nuclear magnetic resonance spectroscopy
alpha-aminopyridine
Proton Magnetic Resonance Spectroscopy

Keywords

  • Bovine
  • Milk fat globule membrane
  • N-linked sugar chain
  • PAS-7 glycoprotein

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biotechnology
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry

Cite this

Structures of the N-linked sugar chains in PAS-7 glycoprotein sharing the same protein core with PAS-6 glycoprotein from the bovine milk fat globule membrane. / Seok, Jin Seok; Shimoda, Michiko; Azuma, Norihiro; Kanno, Choemon.

In: Bioscience, Biotechnology and Biochemistry, Vol. 65, No. 4, 01.04.2001, p. 901-912.

Research output: Contribution to journalArticle

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abstract = "Glycoproteins PAS-6 (50 kDa) and -7 (47 kDa) from the bovine milk fat globule membrane share a common protein core but differ in their carbohydrate moiety. We here analyzed and proposed the structures of the N-linked sugar chains of PAS-7. The N-linked sugar chains were liberated from PAS-7 by hydrazinolysis and, after modifying the reducing ends with 2-aminopyridine (PA), were separated into one neutral (7N, 55{\%}) and two acidic (7M, mono-, 43{\%}; 7D, di-, 2{\%}) sugar chain groups. The latter were converted into neutral groups (7MN and 7DN) by sialidase digestion. 7N was finally separated into 5 chains (7N1A, 7N1B-1, 7N1B-2, 7N2A, and 7N2B), and 7MN and 7DN were separated into 3 (7MN1, 7MN2, and 7MN3) and 2 (7DN1 and 7DN2) chains, respectively. The structure of each of these PA-neutral sugar chains was determined by sugar analysis, sequential exoglycosidase digestion, partial acetolysis, and 1H-NMR spectroscopy. The results show that the 10 sugar chains were of the biantennary complex type with and without fucose. The structure of 7N2A, one of the major sugar chains, was proposed as; Galβ1-4GlcNAcβ1-2Manα1↘ 63 Manβ1-4GlcNAcβ1-4GlcNAc-PA Galβ1-4GlcNAcβ1-2Manα1↗ A structural comparison between PAS-6 and -7 indicated that, although they shared the same protein core, their sugar moiety was markedly different, involving the existence of a different pathway during the post-transcriptional modification.",
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T1 - Structures of the N-linked sugar chains in PAS-7 glycoprotein sharing the same protein core with PAS-6 glycoprotein from the bovine milk fat globule membrane

AU - Seok, Jin Seok

AU - Shimoda, Michiko

AU - Azuma, Norihiro

AU - Kanno, Choemon

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N2 - Glycoproteins PAS-6 (50 kDa) and -7 (47 kDa) from the bovine milk fat globule membrane share a common protein core but differ in their carbohydrate moiety. We here analyzed and proposed the structures of the N-linked sugar chains of PAS-7. The N-linked sugar chains were liberated from PAS-7 by hydrazinolysis and, after modifying the reducing ends with 2-aminopyridine (PA), were separated into one neutral (7N, 55%) and two acidic (7M, mono-, 43%; 7D, di-, 2%) sugar chain groups. The latter were converted into neutral groups (7MN and 7DN) by sialidase digestion. 7N was finally separated into 5 chains (7N1A, 7N1B-1, 7N1B-2, 7N2A, and 7N2B), and 7MN and 7DN were separated into 3 (7MN1, 7MN2, and 7MN3) and 2 (7DN1 and 7DN2) chains, respectively. The structure of each of these PA-neutral sugar chains was determined by sugar analysis, sequential exoglycosidase digestion, partial acetolysis, and 1H-NMR spectroscopy. The results show that the 10 sugar chains were of the biantennary complex type with and without fucose. The structure of 7N2A, one of the major sugar chains, was proposed as; Galβ1-4GlcNAcβ1-2Manα1↘ 63 Manβ1-4GlcNAcβ1-4GlcNAc-PA Galβ1-4GlcNAcβ1-2Manα1↗ A structural comparison between PAS-6 and -7 indicated that, although they shared the same protein core, their sugar moiety was markedly different, involving the existence of a different pathway during the post-transcriptional modification.

AB - Glycoproteins PAS-6 (50 kDa) and -7 (47 kDa) from the bovine milk fat globule membrane share a common protein core but differ in their carbohydrate moiety. We here analyzed and proposed the structures of the N-linked sugar chains of PAS-7. The N-linked sugar chains were liberated from PAS-7 by hydrazinolysis and, after modifying the reducing ends with 2-aminopyridine (PA), were separated into one neutral (7N, 55%) and two acidic (7M, mono-, 43%; 7D, di-, 2%) sugar chain groups. The latter were converted into neutral groups (7MN and 7DN) by sialidase digestion. 7N was finally separated into 5 chains (7N1A, 7N1B-1, 7N1B-2, 7N2A, and 7N2B), and 7MN and 7DN were separated into 3 (7MN1, 7MN2, and 7MN3) and 2 (7DN1 and 7DN2) chains, respectively. The structure of each of these PA-neutral sugar chains was determined by sugar analysis, sequential exoglycosidase digestion, partial acetolysis, and 1H-NMR spectroscopy. The results show that the 10 sugar chains were of the biantennary complex type with and without fucose. The structure of 7N2A, one of the major sugar chains, was proposed as; Galβ1-4GlcNAcβ1-2Manα1↘ 63 Manβ1-4GlcNAcβ1-4GlcNAc-PA Galβ1-4GlcNAcβ1-2Manα1↗ A structural comparison between PAS-6 and -7 indicated that, although they shared the same protein core, their sugar moiety was markedly different, involving the existence of a different pathway during the post-transcriptional modification.

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