Article

Characterization of two novel nodule-enhanced α-type carbonic anhydrases from Lotus japonicus.

Laboratory of Molecular Biology, Department of Agricultural Biotechnology, Agricultural University of Athens, Iera Odos 75, 11855, Athens, Greece.
Biochimica et Biophysica Acta (impact factor: 4.66). 01/2011; 1814(4):496-504. DOI:10.1016/j.bbapap.2011.01.007 pp.496-504
Source: PubMed

ABSTRACT Two cDNA clones coding for α-type carbonic anhydrases (CA; EC 4.2.1.1) in the nitrogen-fixing nodules of the model legume Lotus japonicus were identified. Functionality of the full-length proteins was confirmed by heterologous expression in Escherichia coli and purification of the encoded polypeptides. The developmental expression pattern of LjCAA1 and LjCAA2 revealed that both genes code for nodule enhanced carbonic anhydrase isoforms, which are induced early during nodule development. The genes were slightly to moderately down-regulated in ineffective nodules formed by mutant Mesorhizobium loti strains, indicating that these genes may also be involved in biochemical and physiological processes not directly linked to nitrogen fixation/assimilation. The spatial expression profiling revealed that both genes were expressed in nodule inner cortical cells, vascular bundles and central tissue. These results are discussed in the context of the possible roles of CA in nodule carbon dioxide (CO(2)) metabolism.

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Keywords

biochemical
 
carbonic anhydrase isoforms
 
central tissue
 
developmental expression pattern
 
encoded polypeptides
 
Escherichia coli
 
full-length proteins
 
genes
 
genes code
 
model legume Lotus japonicus
 
moderately down-regulated
 
mutant Mesorhizobium loti strains
 
nitrogen fixation/assimilation
 
nodule carbon dioxide
 
nodule inner cortical cells
 
physiological processes
 
possible roles
 
vascular bundles
 
α-type carbonic anhydrases