title: An external delta-carbonic anhydrase in a free-living marine dinoflagellate may circumvent diffusion-limited carbon acquisition.
abstract: The oceans globally constitute an important sink for carbon dioxide (CO(2)) due to phytoplankton photosynthesis. However, the marine environment imposes serious restraints to carbon fixation. First, the equilibrium between CO(2) and bicarbonate (HCO(3)(-)) is pH dependent, and, in normal, slightly alkaline seawater, [CO(2)] is typically low (approximately 10 mum). Second, the rate of CO(2) diffusion in seawater is slow, so, for any cells unable to take up bicarbonate efficiently, photosynthesis could become carbon limited due to depletion of CO(2) from their immediate vicinity. This may be especially problematic for those dinoflagellates using a form II Rubisco because this form is less oxygen tolerant than the usually found form I enzyme. We have identified a carbonic anhydrase (CA) from the free-living marine dinoflagellate Lingulodinium polyedrum that appears to play a role in carbon acquisition. This CA shares 60% sequence identity with delta-class CAs, isoforms so far found only in marine algae. Immunoelectron microscopy indicates that this enzyme is associated exclusively with the plasma membrane. Furthermore, this enzyme appears to be exposed to the external medium as determined by whole-cell CA assays and vectorial labeling of cell surface proteins with (125)I. The fixation of (14)CO(2) is strongly pH dependent, suggesting preferential uptake of CO(2) rather than HCO(3)(-), and photosynthetic rates decrease in the presence of 1 mm acetazolamide, a non-membrane-permeable CA inhibitor. This constitutes the first CA identified in the dinoflagellates, and, taken together, our results suggest that this enzyme may help to increase CO(2) availability at the cell surface.
authors.affiliation: Institut de Recherche en Biologie Végétale, Département de Sciences Biologiques, Université de Montréal, Montreal, Quebec, Canada H1X 2B2.
authors.forename: Mathieu
authors.initials: M
authors.lastname: Lapointe
authors.forename: Tyler D B
authors.initials: TD
authors.lastname: Mackenzie
authors.forename: David
authors.initials: D
authors.lastname: Morse
mesh_terms: D000086:Acetazolamide / Q000494:pharmacology; D000818:Animals; D001639:Bicarbonates / Q000378:metabolism; D002245:Carbon Dioxide / Q000378:metabolism; D002257:Carbonic Anhydrase Inhibitors / Q000494:pharmacology; D002256:Carbonic Anhydrases / Q000235:genetics / Q000378:metabolism*; D002462:Cell Membrane / Q000201:enzymology*; D004141:Dinoflagellida / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D007527:Isoenzymes / Q000378:metabolism; D010788:Photosynthesis / Q000187:drug effects
chemical_list: D001639:Bicarbonates; D002257:Carbonic Anhydrase Inhibitors; D007527:Isoenzymes; D002245:Carbon Dioxide; D002256:Carbonic Anhydrases; D000086:Acetazolamide