title: Exploring the membrane proteome of the diazotropic cyanobacterium Anabaena PCC7120 through gel-based proteomics and in silico approaches.
abstract: This paper focuses on the gel-based membrane proteomics from diazotrophic cyanobacterium Anabaena PCC7120 by modifying the protocol of Hall et al. [1]. The bioinformatic analysis revealed that 59 (29 integral, 30 peripheral) of the 67 proteins identified were membrane proteins. Of the 29 integral proteins, except Alr0834, the remaining 28 contained 1-12 transmembrane helices. Sixteen integral proteins harboring signal peptides (Sec/TAT/LipoP) suggest that protein targeting in Anabaena involves both sec-dependent and sec-independent pathways. While majority of photosynthesis and respiration proteins (21 of 24) were confined to broad pH gradient the hypothetical and unknown (12 of 13), and cell envelope proteins (3 of 3) preferred the narrow pH range. Of the 5 transporters and binding proteins, Na(+)/H(+)-exchanging protein and Alr2372 were present in broad, pstS1 and cmpD in narrow and cmpA was common to both pH ranges. The distribution of proteins across pH gradient, thus clearly indicates the functional and structural diversity in membrane proteome of Anabaena. It requires mention that protochlorophyllide oxido-reductase, Na(+)/H(+)-exchanging protein, All1355, Alr2055, Alr3514, Alr2903 and Alr2751 were new entries to the 2DE membrane protein profile of Anabaena. This study demonstrates suitability of the modified protocol for the study of membrane protein from filamentous cyanobacteria.
  
  
  CONCLUSIONS
  Anabaena sp. PCC7120 is used as a model organism due to its agriculture significance as biofertilizer, close resemblance with higher plant chloroplast and availability of full genome sequence. Although cytosolic proteome has been explored a lot membrane proteins are still understudied as they are notoriously difficult to display using 2-D technology. Identification and characterization of these proteins is therefore required to elucidate and understand cellular mechanisms. The purpose of this study was to develop a protocol suitable for membrane protein extraction from Anabaena. Additionally, by homology comparison or domain assignment a possible function could be ascribed to novel uncharacterized proteins which will serve as a useful reference for further detailed studies of membrane system in filamentous cyanobacteria. Resolution of membrane proteins ranging from least (single transmembrane helix) to highly hydrophobic (several transmembrane helices) one on 2D gels recommends the gel based approach for identification of membrane proteomics from filamentous cyanobacteria. This article is part of a Special Issue entitled: Proteomics in India.
authors.affiliation: Molecular Biology Section, Centre of Advanced Study in Botany, Banaras Hindu University, Varanasi 221005, India.
authors.forename: Sonia
authors.initials: S
authors.lastname: Sen
authors.affiliation: Molecular Biology Section, Centre of Advanced Study in Botany, Banaras Hindu University, Varanasi 221005, India.
authors.forename: Chhavi
authors.initials: C
authors.lastname: Agrawal
authors.affiliation: Department of Botany, Punjab University, Sector 14, Chandigarh 160014, India.
authors.forename: Yogesh
authors.initials: Y
authors.lastname: Mishra
authors.affiliation: Molecular Biology Section, Centre of Advanced Study in Botany, Banaras Hindu University, Varanasi 221005, India.
authors.forename: Shweta
authors.initials: S
authors.lastname: Rai
authors.affiliation: Molecular Biology Section, Centre of Advanced Study in Botany, Banaras Hindu University, Varanasi 221005, India.
authors.forename: Antra
authors.initials: A
authors.lastname: Chatterjee
authors.affiliation: Molecular Biology Section, Centre of Advanced Study in Botany, Banaras Hindu University, Varanasi 221005, India.
authors.forename: Shivam
authors.initials: S
authors.lastname: Yadav
authors.affiliation: Molecular Biology Section, Centre of Advanced Study in Botany, Banaras Hindu University, Varanasi 221005, India.
authors.forename: Shilpi
authors.initials: S
authors.lastname: Singh
authors.affiliation: Molecular Biology Section, Centre of Advanced Study in Botany, Banaras Hindu University, Varanasi 221005, India. Electronic address: lcrbhu15@gmail.com.
authors.forename: L C
authors.initials: LC
authors.lastname: Rai
mesh_terms: D017033:Anabaena / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism*; D002352:Carrier Proteins / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics
chemical_list: D001426:Bacterial Proteins; D002352:Carrier Proteins; D008565:Membrane Proteins; D020543:Proteome
keywords: Broad and narrow pH gradients; Filamentous cyanobacteria; Gel-based membrane proteomics; Membrane proteins; Two-dimensional (2D) gel electrophoresis