Development of a sample preparation method for fungal proteomics

Motoyuki Shimizu, Hiroyuki Wariishi

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Since filamentous fungi including basidiomycetous fungi possess an exceptionally robust cell wall as in microorganisms, effective extraction of intracellular proteins is a key step for fungal proteomic studies. To overcome the experimental obstacle caused by cell walls, we utilized fungal protoplasts, prepared from the brown-rot basidiomycete, Tyromyces palustris. The amount and quality of proteins extracted from the protoplast cells were much higher than that from the mycelial cells. Quantitative comparisons of proteome maps prepared from mycelial and protoplast cells indicated protein spots with a wider range of molecular weights and pIs in the protoplast sample. Furthermore, no streaking or tailing was observed in the protoplasts, suggesting that effective extraction of intracellular proteins from protoplasts might help suppress degradation of proteins during this process. In addition to the efficiency of protein extraction, simple and efficient subcellular fractionation was also achieved using protoplast cells.

Original languageEnglish
Pages (from-to)17-22
Number of pages6
JournalFEMS microbiology letters
Volume247
Issue number1
DOIs
Publication statusPublished - Jun 1 2005

Fingerprint

Protoplasts
Proteomics
Proteins
Cell Wall
Fungi
Basidiomycota
Proteome
Proteolysis
Molecular Weight

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Molecular Biology
  • Genetics

Cite this

Development of a sample preparation method for fungal proteomics. / Shimizu, Motoyuki; Wariishi, Hiroyuki.

In: FEMS microbiology letters, Vol. 247, No. 1, 01.06.2005, p. 17-22.

Research output: Contribution to journalArticle

@article{9325d08e85a449cea45f3639364975fa,
title = "Development of a sample preparation method for fungal proteomics",
abstract = "Since filamentous fungi including basidiomycetous fungi possess an exceptionally robust cell wall as in microorganisms, effective extraction of intracellular proteins is a key step for fungal proteomic studies. To overcome the experimental obstacle caused by cell walls, we utilized fungal protoplasts, prepared from the brown-rot basidiomycete, Tyromyces palustris. The amount and quality of proteins extracted from the protoplast cells were much higher than that from the mycelial cells. Quantitative comparisons of proteome maps prepared from mycelial and protoplast cells indicated protein spots with a wider range of molecular weights and pIs in the protoplast sample. Furthermore, no streaking or tailing was observed in the protoplasts, suggesting that effective extraction of intracellular proteins from protoplasts might help suppress degradation of proteins during this process. In addition to the efficiency of protein extraction, simple and efficient subcellular fractionation was also achieved using protoplast cells.",
author = "Motoyuki Shimizu and Hiroyuki Wariishi",
year = "2005",
month = "6",
day = "1",
doi = "10.1016/j.femsle.2005.04.021",
language = "English",
volume = "247",
pages = "17--22",
journal = "FEMS Microbiology Letters",
issn = "0378-1097",
publisher = "Wiley-Blackwell",
number = "1",

}

TY - JOUR

T1 - Development of a sample preparation method for fungal proteomics

AU - Shimizu, Motoyuki

AU - Wariishi, Hiroyuki

PY - 2005/6/1

Y1 - 2005/6/1

N2 - Since filamentous fungi including basidiomycetous fungi possess an exceptionally robust cell wall as in microorganisms, effective extraction of intracellular proteins is a key step for fungal proteomic studies. To overcome the experimental obstacle caused by cell walls, we utilized fungal protoplasts, prepared from the brown-rot basidiomycete, Tyromyces palustris. The amount and quality of proteins extracted from the protoplast cells were much higher than that from the mycelial cells. Quantitative comparisons of proteome maps prepared from mycelial and protoplast cells indicated protein spots with a wider range of molecular weights and pIs in the protoplast sample. Furthermore, no streaking or tailing was observed in the protoplasts, suggesting that effective extraction of intracellular proteins from protoplasts might help suppress degradation of proteins during this process. In addition to the efficiency of protein extraction, simple and efficient subcellular fractionation was also achieved using protoplast cells.

AB - Since filamentous fungi including basidiomycetous fungi possess an exceptionally robust cell wall as in microorganisms, effective extraction of intracellular proteins is a key step for fungal proteomic studies. To overcome the experimental obstacle caused by cell walls, we utilized fungal protoplasts, prepared from the brown-rot basidiomycete, Tyromyces palustris. The amount and quality of proteins extracted from the protoplast cells were much higher than that from the mycelial cells. Quantitative comparisons of proteome maps prepared from mycelial and protoplast cells indicated protein spots with a wider range of molecular weights and pIs in the protoplast sample. Furthermore, no streaking or tailing was observed in the protoplasts, suggesting that effective extraction of intracellular proteins from protoplasts might help suppress degradation of proteins during this process. In addition to the efficiency of protein extraction, simple and efficient subcellular fractionation was also achieved using protoplast cells.

UR - http://www.scopus.com/inward/record.url?scp=20344376178&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=20344376178&partnerID=8YFLogxK

U2 - 10.1016/j.femsle.2005.04.021

DO - 10.1016/j.femsle.2005.04.021

M3 - Article

C2 - 15927743

AN - SCOPUS:20344376178

VL - 247

SP - 17

EP - 22

JO - FEMS Microbiology Letters

JF - FEMS Microbiology Letters

SN - 0378-1097

IS - 1

ER -