Amaya Blanco-Rivero

• PhD
• Researcher at Complutense University of Madrid

Introduction Microorganisms colonize a wide range of natural and artificial environments. Even though most of them are unculturable in laboratory conditions, some ecosystems are ideal niches for bioprospecting extremophiles with unique properties. Up today, there are few reports concerning microbial communities found on solar panels, a widespread,...

Chlamydomonas reinhardtii is a green unicellular eukaryotic model organism for studying biological and biotechnological relevant questions. The availability of large genomic resources and the growing interest for C. reinhardtii as an emerging cell factory for the industrial production of biopharmaceuticals requires an in-depth analysis of protein N...

Background: Cah3 is the only carbonic anhydrase (CA) isoform located in the thylakoid lumen of Chlamydomonas reinhardtii. Previous studies demonstrated its association with the donor side of the photosystem II (PSII) where it is required for the optimal function of the water oxidizing complex. However this enzyme has also been frequently proposed...

Association of Cah3 polypeptide with PSII core complexes from high- and low-CO2-grown C. reinhardtii cells. Immunoblot analysis of thylakoid membranes (Thy), BBY preparations (BBY), and PSII core complexes (Core) from cells of C. reinhardtii with antibodies raised against the over-expressed Cah3 polypeptide (Cah3) and D1 protein of PSII. The lanes...

Localization of HA-tagged CAH1 by immunogold labelling and electron microscopy. (A and B) Transmission Electron Microscopy (TEM) images of wt Arabidopsis suspension culture cells. (A) Detailed image of a chloroplast. (B) General view of the cell with ER, Golgi (G), cell wall (CW), vacuole (V), and chloroplast (C). The immunogold (IG) labelling over...

CAH1 contains an intramolecular disulphide bridge. (A) ClustalW2 [47] protein sequence analysis of Arabidopsis α-CA homologues. Cysteine residues at positions 27 and 191 (red) in CAH1 sequence are conserved (*) in seven of the eight homologues described by Fabre et al. [4]. CA8 is substantially larger than CA1-7, possibly indicating that this gene...

TargetP analysis of Arabidopsis α-type CAs. TargetP [8] analysis of Arabidopsis α-type CAs protein sequences described by Fabre et al. [4]. Seven of the eight homologues have an N-terminal signal sequence (SP) for the ER. (TIF)

Cycloheximide treatment results in a higher proportion of Endo H resistant HC. (A) Protoplasts from Arabidopsis cell suspension culture were transiently transfected with HA-tagged wt CAH1 (HC). Prior to protein extraction, protoplasts were either incubated in presence or absence of the protein biosynthesis inhibitor cycloheximide (CHX) for 8 h. Ext...

HA-tagged CAH1 is glycosylated in plant cells, harbouring four or five N-glycans. Migration patterns of HA-tagged CAH1 forms in protoplasts of Arabidopsis suspension culture cells (A) and Arabidopsis mesophyll cells (B) expressing wt (HC) or mutant CAH1. Antibody specificity was verified using water-transfected protoplasts (−). (A) Protoplasts from...

Enrichment of ER marker in microsome fraction. Western blot using antibodies against the ER localized protein BiP to verify that microsome fractions in Figure 4a and b were intact. S and I, soluble supernatant and insoluble microsome pellet, respectively. (TIF)

Nomenclature of constructs used to transfect plant cells and protoplasts. (DOC)

The Arabidopsis CAH1 alpha-type carbonic anhydrase is one of the few plant proteins known to be targeted to the chloroplast through the secretory pathway. CAH1 is post-translationally modified at several residues by the attachment of N-glycans, resulting in a mature protein harbouring complex-type glycans. The reason of why trafficking through this...

Anabaena sp. PCC7120 contains a gene, mrpA (all1838), which forms part of a seven gene-cluster (all1843–all1837) with significant sequence similarity to bacterial operons that putatively code for a multicomponent cation/proton antiporter involved in alkaline pH adaptation and salt resistance. We previously showed that growth and photosynthesis were...

Nowadays, cellular bioenergetics has become a central issue of investigation in cancer biology. Recently, the metabolic activity of the cancer cell has been shown to correlate with a proteomic index that informs of the relative mitochondrial activity of the cell. Within this new field of investigation, we report herein the production and characteri...

Mitochondrial research has experienced a considerable boost during the last decade because organelle malfunctioning is in the genesis and/or progression of a vast array of human pathologies including cancer. The renaissance of mitochondria in the cancer field has been promoted by two main facts: (1) the molecular and functional integration of mitoc...

Mapping of in vivo protein phosphorylation sites in photosynthetic membranes of the green alga Chlamydomonas reinhardtii revealed that the major environmentally dependent changes in phosphorylation are clustered at the interface between the photosystem II (PSII) core and its light-harvesting antennae (LHCII). The photosynthetic membranes that were...

Acclimation of the green alga Chlamydomonas reinhardtii to limiting environmental CO2 induced specific protein phosphorylation at the surface of photosynthetic thylakoid membranes. Four phosphopeptides were identified and sequenced by nanospray quadrupole TOF MS from the cells acclimating to limiting CO2. One phosphopeptide originated from a protei...

A genomic analysis of putative penicillin-binding proteins (PBPs) that are involved in the synthesis of the peptidoglycan layer of the cell wall and are encoded in 12 cyanobacterial genomes was performed in order to help elucidate the role(s) of these proteins in peptidoglycan synthesis, especially during cyanobacterial cellular differentiation. Th...

Transposon mutagenesis of Anabaena sp. PCC7120 led to the isolation of a mutant strain, PHB11, which grew poorly at pH values above 10. The mutant strain exhibited pronounced Na+ sensitivity; this sensitivity was higher under basic conditions. Mutant PHB11 also showed an inhibition of photosynthesis that was much more pronounced at alkaline pH. Rec...

Transposon mutagenesis of Anabaena sp. strain PCC7120 led to the isolation of a mutant strain, SNa1, which is unable to fix nitrogen aerobically but is perfectly able to grow with combined nitrogen (i.e., nitrate). Reconstruction of the transposon mutation of SNa1 in the wild-type strain reproduced the phenotype of the original mutant. The transpos...