Determination and mutational analysis of the phosphorylation site in the hypusine-containing protein Hyp2p

Hannelore Klier; Thorsten Wöhl; Christoph Eckerskorn; Viktor Magdolen; Friedrich Lottspeich

doi:10.1016/0014-5793(93)80712-4

Determination and mutational analysis of the phosphorylation site in the hypusine-containing protein Hyp2p

Hannelore Klier
, Thorsten Wöhl
, Christoph Eckerskorn
, Viktor Magdolen
, Friedrich Lottspeich

Department of Gynecology

Max Planck Institute of Biochemistry

Research output: Contribution to journal › Article › peer-review

23 Scopus citations

Abstract

Electrospray mass spectrometry of the purified isoforms of the hypusine-containing protein of Saccharomyces cerevisiae Hyp2p suggested a phosphorylation of the acidic isoform, which was confirmed by phosphatase treatment. The phosphorylation site was mapped to the N-acetylated serine residue in position no. 1 by mass spectrometric analysis of enzymatic fragments. Mutation of this serine residue gives rise to only the basic isoform, confirming our protein chemical data. As this mutation has no effect on cell viability or growth rate, the unphosphorylated isoform is sufficient to exert the essential in vivo function of Hyp2p.

Original language	English
Pages (from-to)	360-364
Number of pages	5
Journal	FEBS Letters
Volume	334
Issue number	3
DOIs	https://doi.org/10.1016/0014-5793(93)80712-4
State	Published - 22 Nov 1993

Keywords

Acetylation
Amino terminus
Hyp1p, gene product of HYP1 = ANB 1 = TIF51B
Hyp2p, gene product of HYP2 = TIF51A
IEF, isoelectric focussing
Isoform
Phosphorylation
SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
Yeast abr]HPLC, high-performance liquid chromatography
eIF-5A
hyp2, disrupted HYP2 gene
pI, isoelectric point

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10.1016/0014-5793(93)80712-4

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title = "Determination and mutational analysis of the phosphorylation site in the hypusine-containing protein Hyp2p",

abstract = "Electrospray mass spectrometry of the purified isoforms of the hypusine-containing protein of Saccharomyces cerevisiae Hyp2p suggested a phosphorylation of the acidic isoform, which was confirmed by phosphatase treatment. The phosphorylation site was mapped to the N-acetylated serine residue in position no. 1 by mass spectrometric analysis of enzymatic fragments. Mutation of this serine residue gives rise to only the basic isoform, confirming our protein chemical data. As this mutation has no effect on cell viability or growth rate, the unphosphorylated isoform is sufficient to exert the essential in vivo function of Hyp2p.",

keywords = "Acetylation, Amino terminus, Hyp1p, gene product of HYP1 = ANB 1 = TIF51B, Hyp2p, gene product of HYP2 = TIF51A, IEF, isoelectric focussing, Isoform, Phosphorylation, SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis., Yeast abr]HPLC, high-performance liquid chromatography, eIF-5A, hyp2, disrupted HYP2 gene, pI, isoelectric point",

author = "Hannelore Klier and Thorsten W{\"o}hl and Christoph Eckerskorn and Viktor Magdolen and Friedrich Lottspeich",

year = "1993",

month = nov,

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doi = "10.1016/0014-5793(93)80712-4",

language = "English",

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journal = "FEBS Letters",

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TY - JOUR

T1 - Determination and mutational analysis of the phosphorylation site in the hypusine-containing protein Hyp2p

AU - Klier, Hannelore

AU - Wöhl, Thorsten

AU - Eckerskorn, Christoph

AU - Magdolen, Viktor

AU - Lottspeich, Friedrich

PY - 1993/11/22

Y1 - 1993/11/22

N2 - Electrospray mass spectrometry of the purified isoforms of the hypusine-containing protein of Saccharomyces cerevisiae Hyp2p suggested a phosphorylation of the acidic isoform, which was confirmed by phosphatase treatment. The phosphorylation site was mapped to the N-acetylated serine residue in position no. 1 by mass spectrometric analysis of enzymatic fragments. Mutation of this serine residue gives rise to only the basic isoform, confirming our protein chemical data. As this mutation has no effect on cell viability or growth rate, the unphosphorylated isoform is sufficient to exert the essential in vivo function of Hyp2p.

AB - Electrospray mass spectrometry of the purified isoforms of the hypusine-containing protein of Saccharomyces cerevisiae Hyp2p suggested a phosphorylation of the acidic isoform, which was confirmed by phosphatase treatment. The phosphorylation site was mapped to the N-acetylated serine residue in position no. 1 by mass spectrometric analysis of enzymatic fragments. Mutation of this serine residue gives rise to only the basic isoform, confirming our protein chemical data. As this mutation has no effect on cell viability or growth rate, the unphosphorylated isoform is sufficient to exert the essential in vivo function of Hyp2p.

KW - Acetylation

KW - Amino terminus

KW - Hyp1p, gene product of HYP1 = ANB 1 = TIF51B

KW - Hyp2p, gene product of HYP2 = TIF51A

KW - IEF, isoelectric focussing

KW - Isoform

KW - Phosphorylation

KW - SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

KW - Yeast abr]HPLC, high-performance liquid chromatography

KW - eIF-5A

KW - hyp2, disrupted HYP2 gene

KW - pI, isoelectric point

UR - https://www.scopus.com/pages/publications/0027426296

U2 - 10.1016/0014-5793(93)80712-4

DO - 10.1016/0014-5793(93)80712-4

M3 - Article

C2 - 8243648

AN - SCOPUS:0027426296

SN - 0014-5793

VL - 334

SP - 360

EP - 364

JO - FEBS Letters

JF - FEBS Letters

IS - 3

ER -

Determination and mutational analysis of the phosphorylation site in the hypusine-containing protein Hyp2p

Abstract

Keywords

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