NO-degradation by alfalfa class 1 hemoglobin (Mhb1): A possible link to PR-1a gene expression in Mhb1-overproducing tobacco plants

Csaba Seregélyes, Abir U. Igamberdiev, Anna Maassen, Jacek Hennig, D. Dudits, Robert D. Hill

Research output: Contribution to journalArticle

52 Citations (Scopus)

Abstract

Tobacco plants overproducing alfalfa class 1 hemoglobin (HOT plants) have been shown to have reduced necrotic symptom development. Here, we show that this altered pathogenic response is linked to a significant increase in the nitric oxide (NO)-affected pathogenesis-related (PR-1a) transcript accumulation in the transgenic plants. Homogenates of HOT transgenic seedlings were also found to have higher NO-scavenging activity than non-transformed ones. The NO-scavenging properties of recombinant alfalfa class1 hemoglobin have been examined. Recombinant Mhb1 (rMhb1) was produced in bacteria and purified using polyethylene glycol (10-25%) fractionation, chromatography on DEAE-Sephacel, and Phenyl Superose columns. After the final purification step, the obtained preparations were near homogeneous and had a molecular weight of 44 kDa determined by size-exclusion chromatography and 23 kDa by SDS-PAGE, indicating that rMhb1 is a dimer. The protein participated in NO-degradation activity with NAD(P)H as a cofactor. After ion-exchange columns, addition of FAD was necessary for exhibiting maximal NO-degradation activity. The NAD(P)H-dependent NO-scavenging activity of rMhb1, which is similar to that of barley hemoglobin, supports a conclusion that both monocot and dicot class 1 hemoglobins can affect cellular NO levels by scavenging NO formed during hypoxia, pathogen attack and other stresses.

Original languageEnglish
Pages (from-to)61-66
Number of pages6
JournalFEBS Letters
Volume571
Issue number1-3
DOIs
Publication statusPublished - Jul 30 2004

Fingerprint

Medicago sativa
Tobacco
Glycosylated Hemoglobin A
Gene expression
Nitric Oxide
Hemoglobins
Gene Expression
Degradation
Scavenging
NAD
Flavin-Adenine Dinucleotide
Size exclusion chromatography
Ion Exchange
Genetically Modified Plants
Distillation columns
Pathogens
Fractionation
Chromatography
Seedlings
Dimers

Keywords

  • Class1 hemoglobin
  • Hexacoordinate
  • Nitric oxide
  • Pathogen
  • PR-1a
  • Purification

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

NO-degradation by alfalfa class 1 hemoglobin (Mhb1) : A possible link to PR-1a gene expression in Mhb1-overproducing tobacco plants. / Seregélyes, Csaba; Igamberdiev, Abir U.; Maassen, Anna; Hennig, Jacek; Dudits, D.; Hill, Robert D.

In: FEBS Letters, Vol. 571, No. 1-3, 30.07.2004, p. 61-66.

Research output: Contribution to journalArticle

Seregélyes, Csaba ; Igamberdiev, Abir U. ; Maassen, Anna ; Hennig, Jacek ; Dudits, D. ; Hill, Robert D. / NO-degradation by alfalfa class 1 hemoglobin (Mhb1) : A possible link to PR-1a gene expression in Mhb1-overproducing tobacco plants. In: FEBS Letters. 2004 ; Vol. 571, No. 1-3. pp. 61-66.
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