Reversed-phase high-performance liquid chromatographic separation of synthetic phosphopeptide isomers

Laszlo Otvos, Ibrahim A. Tangoren, Krzysztof Wroblewski, M. Hollósi, Virginia M Y Lee

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Selectively phosphorylated synthetic peptides corresponding to the human neurofilament protein middle-sized subunit, H-Lys-Ser-Pro-Val-Pro-Lys-Ser-Pro-Val-Glu-Glu-Lys-Gly-Oh, and its analogues were separated by reversed-phase high-performance liquid chromatography of mixtures consisting of the non-phosphorylated, the diphosphorylated and the two different monophosphorylated isomers. Application of the algorithm for the expected retention times to 4-9 amino acid-long peptide fragments revealed the correct elution order of the monophosphorylated isomers. According to circular dichroism studies, this elution order is also compatible with the possibility of induced conformational orientation on the surface of the bonded phase. Chromatographic analysis of the synthetic phosphorylation reaction indicates that the reaction rates of the two structurally different monophosphorylated peptides are similar, which is in contrast to the in vivo site-directed reaction.

Original languageEnglish
Pages (from-to)265-272
Number of pages8
JournalJournal of Chromatography A
Volume512
Issue numberC
DOIs
Publication statusPublished - Jul 20 1990

Fingerprint

Phosphopeptides
Isomers
Chromatographic analysis
Neurofilament Proteins
Peptides
Phosphorylation
Peptide Fragments
High performance liquid chromatography
Liquids
Reverse-Phase Chromatography
Circular Dichroism
Reaction rates
Chromatography
High Pressure Liquid Chromatography
Amino Acids

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Reversed-phase high-performance liquid chromatographic separation of synthetic phosphopeptide isomers. / Otvos, Laszlo; Tangoren, Ibrahim A.; Wroblewski, Krzysztof; Hollósi, M.; Lee, Virginia M Y.

In: Journal of Chromatography A, Vol. 512, No. C, 20.07.1990, p. 265-272.

Research output: Contribution to journalArticle

Otvos, Laszlo ; Tangoren, Ibrahim A. ; Wroblewski, Krzysztof ; Hollósi, M. ; Lee, Virginia M Y. / Reversed-phase high-performance liquid chromatographic separation of synthetic phosphopeptide isomers. In: Journal of Chromatography A. 1990 ; Vol. 512, No. C. pp. 265-272.
@article{6f313d00c3694fe9a1b1243032a40d5a,
title = "Reversed-phase high-performance liquid chromatographic separation of synthetic phosphopeptide isomers",
abstract = "Selectively phosphorylated synthetic peptides corresponding to the human neurofilament protein middle-sized subunit, H-Lys-Ser-Pro-Val-Pro-Lys-Ser-Pro-Val-Glu-Glu-Lys-Gly-Oh, and its analogues were separated by reversed-phase high-performance liquid chromatography of mixtures consisting of the non-phosphorylated, the diphosphorylated and the two different monophosphorylated isomers. Application of the algorithm for the expected retention times to 4-9 amino acid-long peptide fragments revealed the correct elution order of the monophosphorylated isomers. According to circular dichroism studies, this elution order is also compatible with the possibility of induced conformational orientation on the surface of the bonded phase. Chromatographic analysis of the synthetic phosphorylation reaction indicates that the reaction rates of the two structurally different monophosphorylated peptides are similar, which is in contrast to the in vivo site-directed reaction.",
author = "Laszlo Otvos and Tangoren, {Ibrahim A.} and Krzysztof Wroblewski and M. Holl{\'o}si and Lee, {Virginia M Y}",
year = "1990",
month = "7",
day = "20",
doi = "10.1016/S0021-9673(01)89493-0",
language = "English",
volume = "512",
pages = "265--272",
journal = "Journal of Chromatography",
issn = "0021-9673",
publisher = "Elsevier",
number = "C",

}

TY - JOUR

T1 - Reversed-phase high-performance liquid chromatographic separation of synthetic phosphopeptide isomers

AU - Otvos, Laszlo

AU - Tangoren, Ibrahim A.

AU - Wroblewski, Krzysztof

AU - Hollósi, M.

AU - Lee, Virginia M Y

PY - 1990/7/20

Y1 - 1990/7/20

N2 - Selectively phosphorylated synthetic peptides corresponding to the human neurofilament protein middle-sized subunit, H-Lys-Ser-Pro-Val-Pro-Lys-Ser-Pro-Val-Glu-Glu-Lys-Gly-Oh, and its analogues were separated by reversed-phase high-performance liquid chromatography of mixtures consisting of the non-phosphorylated, the diphosphorylated and the two different monophosphorylated isomers. Application of the algorithm for the expected retention times to 4-9 amino acid-long peptide fragments revealed the correct elution order of the monophosphorylated isomers. According to circular dichroism studies, this elution order is also compatible with the possibility of induced conformational orientation on the surface of the bonded phase. Chromatographic analysis of the synthetic phosphorylation reaction indicates that the reaction rates of the two structurally different monophosphorylated peptides are similar, which is in contrast to the in vivo site-directed reaction.

AB - Selectively phosphorylated synthetic peptides corresponding to the human neurofilament protein middle-sized subunit, H-Lys-Ser-Pro-Val-Pro-Lys-Ser-Pro-Val-Glu-Glu-Lys-Gly-Oh, and its analogues were separated by reversed-phase high-performance liquid chromatography of mixtures consisting of the non-phosphorylated, the diphosphorylated and the two different monophosphorylated isomers. Application of the algorithm for the expected retention times to 4-9 amino acid-long peptide fragments revealed the correct elution order of the monophosphorylated isomers. According to circular dichroism studies, this elution order is also compatible with the possibility of induced conformational orientation on the surface of the bonded phase. Chromatographic analysis of the synthetic phosphorylation reaction indicates that the reaction rates of the two structurally different monophosphorylated peptides are similar, which is in contrast to the in vivo site-directed reaction.

UR - http://www.scopus.com/inward/record.url?scp=0025711355&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025711355&partnerID=8YFLogxK

U2 - 10.1016/S0021-9673(01)89493-0

DO - 10.1016/S0021-9673(01)89493-0

M3 - Article

C2 - 2229230

AN - SCOPUS:0025711355

VL - 512

SP - 265

EP - 272

JO - Journal of Chromatography

JF - Journal of Chromatography

SN - 0021-9673

IS - C

ER -