Identification and quantitation of phosphorus metabolites in yeast neutral pH extracts by nuclear magnetic resonance spectroscopy

Research output: Contribution to journalArticleScientificpeer-review

26 Citations (Scopus)

Abstract

31P NMR spectroscopy offers a possibility to obtain a survey of all low-molecular-weight phosphorylated compounds in yeast. The yeast cells have been extracted using chloroform into a neutral aqueous phase. The use of high fields and the neutral pH extracts, which are suitable for NMR analysis, results in well-resolved 31P NMR spectra. Two-dimensional NMR experiments, such as proton-detected heteronuclear single quantum (1H-31P HSQC) and 31P correlation spectroscopy (31P COSY), have been used to assign the resonances. In the phosphomonoester region many of the signals could be assigned to known metabolites in the glycolytic and pentose phosphate pathways, although some signals remain unidentified. Accumulation of ribulose 5-phosphate, xylulose 5-phosphate, and ribose 5-phosphate was observed in a strain lacking transketolase activity when grown in synthetic complete medium. No such accumulation occurred when the cells were grown in yeast- peptone-dextrose medium. Trimetaphosphate (intracellular concentration about 0.2 mM) was detected in both cold methanol-chloroform and perchloric acid extracts.

Original languageEnglish
Pages (from-to)71-79
Number of pages9
JournalAnalytical Biochemistry
Volume272
Issue number1
DOIs
Publication statusPublished - 15 Jul 1999
MoE publication typeA1 Journal article-refereed

Fingerprint

Metabolites
Yeast
Phosphorus
Nuclear magnetic resonance spectroscopy
Magnetic Resonance Spectroscopy
Yeasts
Nuclear magnetic resonance
Chloroform
Transketolase
Pentoses
Pentose Phosphate Pathway
Peptones
Methanol
Protons
Spectrum Analysis
Molecular Weight
Molecular weight
Phosphates
Cells
Spectroscopy

Cite this

@article{925d96b2d0ea4f6184bd37928ae04b92,
title = "Identification and quantitation of phosphorus metabolites in yeast neutral pH extracts by nuclear magnetic resonance spectroscopy",
abstract = "31P NMR spectroscopy offers a possibility to obtain a survey of all low-molecular-weight phosphorylated compounds in yeast. The yeast cells have been extracted using chloroform into a neutral aqueous phase. The use of high fields and the neutral pH extracts, which are suitable for NMR analysis, results in well-resolved 31P NMR spectra. Two-dimensional NMR experiments, such as proton-detected heteronuclear single quantum (1H-31P HSQC) and 31P correlation spectroscopy (31P COSY), have been used to assign the resonances. In the phosphomonoester region many of the signals could be assigned to known metabolites in the glycolytic and pentose phosphate pathways, although some signals remain unidentified. Accumulation of ribulose 5-phosphate, xylulose 5-phosphate, and ribose 5-phosphate was observed in a strain lacking transketolase activity when grown in synthetic complete medium. No such accumulation occurred when the cells were grown in yeast- peptone-dextrose medium. Trimetaphosphate (intracellular concentration about 0.2 mM) was detected in both cold methanol-chloroform and perchloric acid extracts.",
author = "Anita Teleman and Peter Richard and Mervi Toivari and Merja Penttil{\"a}",
year = "1999",
month = "7",
day = "15",
doi = "10.1006/abio.1999.4165",
language = "English",
volume = "272",
pages = "71--79",
journal = "Analytical Biochemistry",
issn = "0003-2697",
publisher = "Academic Press",
number = "1",

}

Identification and quantitation of phosphorus metabolites in yeast neutral pH extracts by nuclear magnetic resonance spectroscopy. / Teleman, Anita; Richard, Peter; Toivari, Mervi; Penttilä, Merja.

In: Analytical Biochemistry, Vol. 272, No. 1, 15.07.1999, p. 71-79.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Identification and quantitation of phosphorus metabolites in yeast neutral pH extracts by nuclear magnetic resonance spectroscopy

AU - Teleman, Anita

AU - Richard, Peter

AU - Toivari, Mervi

AU - Penttilä, Merja

PY - 1999/7/15

Y1 - 1999/7/15

N2 - 31P NMR spectroscopy offers a possibility to obtain a survey of all low-molecular-weight phosphorylated compounds in yeast. The yeast cells have been extracted using chloroform into a neutral aqueous phase. The use of high fields and the neutral pH extracts, which are suitable for NMR analysis, results in well-resolved 31P NMR spectra. Two-dimensional NMR experiments, such as proton-detected heteronuclear single quantum (1H-31P HSQC) and 31P correlation spectroscopy (31P COSY), have been used to assign the resonances. In the phosphomonoester region many of the signals could be assigned to known metabolites in the glycolytic and pentose phosphate pathways, although some signals remain unidentified. Accumulation of ribulose 5-phosphate, xylulose 5-phosphate, and ribose 5-phosphate was observed in a strain lacking transketolase activity when grown in synthetic complete medium. No such accumulation occurred when the cells were grown in yeast- peptone-dextrose medium. Trimetaphosphate (intracellular concentration about 0.2 mM) was detected in both cold methanol-chloroform and perchloric acid extracts.

AB - 31P NMR spectroscopy offers a possibility to obtain a survey of all low-molecular-weight phosphorylated compounds in yeast. The yeast cells have been extracted using chloroform into a neutral aqueous phase. The use of high fields and the neutral pH extracts, which are suitable for NMR analysis, results in well-resolved 31P NMR spectra. Two-dimensional NMR experiments, such as proton-detected heteronuclear single quantum (1H-31P HSQC) and 31P correlation spectroscopy (31P COSY), have been used to assign the resonances. In the phosphomonoester region many of the signals could be assigned to known metabolites in the glycolytic and pentose phosphate pathways, although some signals remain unidentified. Accumulation of ribulose 5-phosphate, xylulose 5-phosphate, and ribose 5-phosphate was observed in a strain lacking transketolase activity when grown in synthetic complete medium. No such accumulation occurred when the cells were grown in yeast- peptone-dextrose medium. Trimetaphosphate (intracellular concentration about 0.2 mM) was detected in both cold methanol-chloroform and perchloric acid extracts.

UR - http://www.scopus.com/inward/record.url?scp=0033566229&partnerID=8YFLogxK

U2 - 10.1006/abio.1999.4165

DO - 10.1006/abio.1999.4165

M3 - Article

C2 - 10405295

AN - SCOPUS:0033566229

VL - 272

SP - 71

EP - 79

JO - Analytical Biochemistry

JF - Analytical Biochemistry

SN - 0003-2697

IS - 1

ER -