Cloning and characterization of a caesalpinoid (Chamaecrista fasciculata) hemoglobin

The structural transition from a nonsymbiotic hemoglobin to a leghemoglobin

Sabarinathan K. Gopalasubramaniam, Frank Kovacs, Fernando Violante-Mota, Paul Twigg, Raúl Arredondo-Peter, Gautam Sarath

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Nonsymbiotic hemoglobins (nsHbs) and leghemoglobins (Lbs) are plant proteins that can reversibly bind O 2 and other ligands. The nsHbs are hexacoordinate and appear to modulate cellular concentrations of NO and maintain energy levels under hypoxic conditions. The Lbs are pentacoordinate and facilitate the diffusion of O 2 to symbiotic bacteroids within legume root nodules. Multiple lines of evidence suggest that all plant Hbs evolved from a common ancestor and that Lbs originated from nsHbs. However, little is known about the structural intermediates that occurred during the evolution of pentacoordinate Lbs from hexacoordinate nsHbs. We have cloned and characterized a Hb (ppHb) from the root nodules of the ancient caesalpinoid legume Chamaecrista fasciculata. Protein sequence, modeling data, and spectral analysis indicated that the properties of ppHb are intermediate between that of nsHb and Lb, suggesting that ppHb resembles a putative ancestral Lb. Predicted structural changes that appear to have occurred during the nsHb to Lb transition were a compaction of the CD-loop and decreased mobility of the distal His inhibiting its ability to coordinate directly with the heme-Fe, leading to a pentacoordinate protein. Other predicted changes include shortening of the N- and C-termini, compaction of the protein into a globular structure, disappearance of positive charges outside the heme pocket and appearance of negative charges in an area located between the N- and C-termini. A major consequence for some of these changes appears to be the decrease in O 2 -affinity of ancestral nsHb, which resulted in the origin of the symbiotic function of Lbs.

Original languageEnglish (US)
Pages (from-to)252-260
Number of pages9
JournalProteins: Structure, Function and Genetics
Volume72
Issue number1
DOIs
StatePublished - Jul 1 2008

Fingerprint

Chamaecrista
Leghemoglobin
Cloning
Organism Cloning
Hemoglobins
Heme
Fabaceae
Compaction
Plant Proteins
Proteins
Spectrum analysis
Electron energy levels
Ligands

Keywords

  • Caesalpinoideae
  • Evolution
  • Function
  • Globin
  • Leguminosae
  • Modeling
  • Origin
  • Spectroscopy
  • Structure
  • Symbiosis

ASJC Scopus subject areas

  • Structural Biology
  • Biochemistry
  • Molecular Biology

Cite this

Cloning and characterization of a caesalpinoid (Chamaecrista fasciculata) hemoglobin : The structural transition from a nonsymbiotic hemoglobin to a leghemoglobin. / Gopalasubramaniam, Sabarinathan K.; Kovacs, Frank; Violante-Mota, Fernando; Twigg, Paul; Arredondo-Peter, Raúl; Sarath, Gautam.

In: Proteins: Structure, Function and Genetics, Vol. 72, No. 1, 01.07.2008, p. 252-260.

Research output: Contribution to journalArticle

Gopalasubramaniam, Sabarinathan K. ; Kovacs, Frank ; Violante-Mota, Fernando ; Twigg, Paul ; Arredondo-Peter, Raúl ; Sarath, Gautam. / Cloning and characterization of a caesalpinoid (Chamaecrista fasciculata) hemoglobin : The structural transition from a nonsymbiotic hemoglobin to a leghemoglobin. In: Proteins: Structure, Function and Genetics. 2008 ; Vol. 72, No. 1. pp. 252-260.
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