Andrea Martinez Bernardini

• Head of Department at Zymvol Biomodeling S.L

In 2012, David Rozzell published an excellent and concise survey of commercial enzyme suppliers. This chapter includes details of companies that provide one or more of the many services focused towards bioprocess development. This survey is primarily aimed at the industrial and academic synthetic chemist who needs to source enzymes or services for...

Timing a switch in tissue integrity In plants, sperm cells travel through the pollen tube as it grows toward the ovule. Successful fertilization depends on the pollen tube rupturing to release the sperm cells (see the Perspective by Stegmann and Zipfel). Ge et al. and Mecchia et al. elucidated the intercellular cross-talk that maintains pollen tube...

New insights into the molecular dialogue between male and female during sexual plant reproduction show that even plant sex does not work without clear communication. Please see related Research article: http://genomebiology.biomedcentral.com/articles/10.1186/s13059-016-0928-x

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...