In Vivo Modifications of the Maize Mitochondrial Small Heat Stress Protein, HSP22

Adrian A. Lund, David M. Rhoads, Anders L. Lund, Ronald Cerny, Thomas E. Elthon

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

A maize (Zea mays L.) small heat shock protein (HSP), HSP22, was previously shown to accumulate to high levels in mitochondria during heat stress. Here we have purified native HSP22 and resolved the protein into three peaks using reverse phase high performance liquid chromatography. Mass spectrometry (MS) of the first two peaks revealed the presence of two HSP22 forms in each peak which differed in mass by 80 daltons (Da), indicative of a monophosphorylation. Phosphorylation of HSP22 by [γ-32P]ATP was also observed in mitochondria labeled in vitro, but not when purified native HSP22 was similarly used, demonstrating that HSP22 does not autophosphorylate, implicating a kinase involvement in vivo. Collisionally induced dissociation tandem MS (CID MS/MS) identified Ser59 as the phosphorylated residue. We have also observed forms of HSP22 that result from alternative intron splicing. The two HSP22 proteins in the first peak were ∼57 Da larger than the two HSP22 proteins in the second peak. MS analysis revealed that the +57-Da forms have an additional Gly residue directly N-terminal of the expected Asp84, which had been converted to an Asn residue. These results are the first demonstrations of phosphorylation and alternative intron splicing of a plant small HSP.

Original languageEnglish (US)
Pages (from-to)29924-29929
Number of pages6
JournalJournal of Biological Chemistry
Volume276
Issue number32
DOIs
StatePublished - Aug 10 2001

Fingerprint

Heat-Shock Proteins
Small Heat-Shock Proteins
Zea mays
Mass spectrometry
Hot Temperature
Alternative Splicing
Tandem Mass Spectrometry
Introns
Mass Spectrometry
Mitochondria
Phosphorylation
Plant Proteins
Proteins
Reverse-Phase Chromatography
Phosphotransferases
Adenosine Triphosphate
High Pressure Liquid Chromatography
High performance liquid chromatography
Demonstrations

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

In Vivo Modifications of the Maize Mitochondrial Small Heat Stress Protein, HSP22. / Lund, Adrian A.; Rhoads, David M.; Lund, Anders L.; Cerny, Ronald; Elthon, Thomas E.

In: Journal of Biological Chemistry, Vol. 276, No. 32, 10.08.2001, p. 29924-29929.

Research output: Contribution to journalArticle

Lund, Adrian A. ; Rhoads, David M. ; Lund, Anders L. ; Cerny, Ronald ; Elthon, Thomas E. / In Vivo Modifications of the Maize Mitochondrial Small Heat Stress Protein, HSP22. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 32. pp. 29924-29929.
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