α-Mannosidase involved in turnover of plant complex type N-glycans in tomato (Lycopersicum esculentum) fruits

Md Anowar Hossain, Kosuke Nakamura, Yoshinobu Kimura

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

In this study, we purified and characterized an α-mannosidase to homogeneity from mature red tomato fruits. Purified α-mannosidase (α-Man LE-1) gave two separate bands, of molecular masses of 70 kDa (L-subunit) and 47 kDa (S-subunit), on SDS-PAGE under non-reducing and reducing conditions. On the other hand, the molecular weight was estimated to be 230 kDa by gel filtration, indicating that α-Man LE-1 functions in a tetrameric structure in plant cells. The N-terminal sequence of the L-subunit and the S-subunit were determined to be L-Y-M-V-Y-M-T-K-Q-G- and X-X-L-E-Q/K-S-F-S-Y-Y respectively. When pyridylaminated N-glycans were used as substrates, α-Man LE-1 showed optimum activity at about pH 6 and at 40°C, and the activity was completely inhibited by both swainsonine and 1-deoxy- mannojirimycin. α-Man LE-1 hydrolyzed the α-mannosidic linkages from both high-mannose type and plant complex type N-glycan, but preferred a truncated plant complex type structure to high-mannose type N-glycans bearing α1-2 mannosyl residues.

Original languageEnglish
Pages (from-to)140-146
Number of pages7
JournalBioscience, Biotechnology and Biochemistry
Volume73
Issue number1
DOIs
Publication statusPublished - 2009

Fingerprint

Mannosidases
Lycopersicon esculentum
Fruits
Polysaccharides
Fruit
Mannose
Bearings (structural)
Swainsonine
Molecular mass
Plant Structures
Plant Cells
Gels
Molecular weight
Gel Chromatography
Polyacrylamide Gel Electrophoresis
Molecular Weight
Substrates

Keywords

  • α-mannosidase
  • Lycopersicon esculentum
  • Plant n-glycan metabolism
  • Tomato fruit ripening

ASJC Scopus subject areas

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

Cite this

α-Mannosidase involved in turnover of plant complex type N-glycans in tomato (Lycopersicum esculentum) fruits. / Hossain, Md Anowar; Nakamura, Kosuke; Kimura, Yoshinobu.

In: Bioscience, Biotechnology and Biochemistry, Vol. 73, No. 1, 2009, p. 140-146.

Research output: Contribution to journalArticle

@article{1e66d2c40ce4407db53ca154ce67d0ae,
title = "α-Mannosidase involved in turnover of plant complex type N-glycans in tomato (Lycopersicum esculentum) fruits",
abstract = "In this study, we purified and characterized an α-mannosidase to homogeneity from mature red tomato fruits. Purified α-mannosidase (α-Man LE-1) gave two separate bands, of molecular masses of 70 kDa (L-subunit) and 47 kDa (S-subunit), on SDS-PAGE under non-reducing and reducing conditions. On the other hand, the molecular weight was estimated to be 230 kDa by gel filtration, indicating that α-Man LE-1 functions in a tetrameric structure in plant cells. The N-terminal sequence of the L-subunit and the S-subunit were determined to be L-Y-M-V-Y-M-T-K-Q-G- and X-X-L-E-Q/K-S-F-S-Y-Y respectively. When pyridylaminated N-glycans were used as substrates, α-Man LE-1 showed optimum activity at about pH 6 and at 40°C, and the activity was completely inhibited by both swainsonine and 1-deoxy- mannojirimycin. α-Man LE-1 hydrolyzed the α-mannosidic linkages from both high-mannose type and plant complex type N-glycan, but preferred a truncated plant complex type structure to high-mannose type N-glycans bearing α1-2 mannosyl residues.",
keywords = "α-mannosidase, Lycopersicon esculentum, Plant n-glycan metabolism, Tomato fruit ripening",
author = "Hossain, {Md Anowar} and Kosuke Nakamura and Yoshinobu Kimura",
year = "2009",
doi = "10.1271/bbb.80561",
language = "English",
volume = "73",
pages = "140--146",
journal = "Bioscience, Biotechnology and Biochemistry",
issn = "0916-8451",
publisher = "Japan Society for Bioscience Biotechnology and Agrochemistry",
number = "1",

}

TY - JOUR

T1 - α-Mannosidase involved in turnover of plant complex type N-glycans in tomato (Lycopersicum esculentum) fruits

AU - Hossain, Md Anowar

AU - Nakamura, Kosuke

AU - Kimura, Yoshinobu

PY - 2009

Y1 - 2009

N2 - In this study, we purified and characterized an α-mannosidase to homogeneity from mature red tomato fruits. Purified α-mannosidase (α-Man LE-1) gave two separate bands, of molecular masses of 70 kDa (L-subunit) and 47 kDa (S-subunit), on SDS-PAGE under non-reducing and reducing conditions. On the other hand, the molecular weight was estimated to be 230 kDa by gel filtration, indicating that α-Man LE-1 functions in a tetrameric structure in plant cells. The N-terminal sequence of the L-subunit and the S-subunit were determined to be L-Y-M-V-Y-M-T-K-Q-G- and X-X-L-E-Q/K-S-F-S-Y-Y respectively. When pyridylaminated N-glycans were used as substrates, α-Man LE-1 showed optimum activity at about pH 6 and at 40°C, and the activity was completely inhibited by both swainsonine and 1-deoxy- mannojirimycin. α-Man LE-1 hydrolyzed the α-mannosidic linkages from both high-mannose type and plant complex type N-glycan, but preferred a truncated plant complex type structure to high-mannose type N-glycans bearing α1-2 mannosyl residues.

AB - In this study, we purified and characterized an α-mannosidase to homogeneity from mature red tomato fruits. Purified α-mannosidase (α-Man LE-1) gave two separate bands, of molecular masses of 70 kDa (L-subunit) and 47 kDa (S-subunit), on SDS-PAGE under non-reducing and reducing conditions. On the other hand, the molecular weight was estimated to be 230 kDa by gel filtration, indicating that α-Man LE-1 functions in a tetrameric structure in plant cells. The N-terminal sequence of the L-subunit and the S-subunit were determined to be L-Y-M-V-Y-M-T-K-Q-G- and X-X-L-E-Q/K-S-F-S-Y-Y respectively. When pyridylaminated N-glycans were used as substrates, α-Man LE-1 showed optimum activity at about pH 6 and at 40°C, and the activity was completely inhibited by both swainsonine and 1-deoxy- mannojirimycin. α-Man LE-1 hydrolyzed the α-mannosidic linkages from both high-mannose type and plant complex type N-glycan, but preferred a truncated plant complex type structure to high-mannose type N-glycans bearing α1-2 mannosyl residues.

KW - α-mannosidase

KW - Lycopersicon esculentum

KW - Plant n-glycan metabolism

KW - Tomato fruit ripening

UR - http://www.scopus.com/inward/record.url?scp=58849111345&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=58849111345&partnerID=8YFLogxK

U2 - 10.1271/bbb.80561

DO - 10.1271/bbb.80561

M3 - Article

C2 - 19129634

AN - SCOPUS:58849111345

VL - 73

SP - 140

EP - 146

JO - Bioscience, Biotechnology and Biochemistry

JF - Bioscience, Biotechnology and Biochemistry

SN - 0916-8451

IS - 1

ER -