Kinetic complexities of triacylglycerol accumulation in developing embryos from Camelina sativa provide evidence for multiple biosynthetic systems

Pollard, Mike; Shachar-Hill, Yair

doi:10.1016/j.jbc.2021.101396

Title: Kinetic complexities of triacylglycerol accumulation in developing embryos from Camelina sativa provide evidence for multiple biosynthetic systems

Journal Article · Sat Jan 01 00:00:00 EST 2022 · Journal of Biological Chemistry

DOI:https://doi.org/10.1016/j.jbc.2021.101396· OSTI ID:1830024

Pollard, Mike;

Quantitative flux maps describing glycerolipid synthesis can be important tools for rational engineering of lipid content and composition in oilseeds. Lipid accumulation in cultured embryos of Camelina sativa is known to mimic that of seeds in terms of rate of lipid synthesis and composition. To assess the kinetic complexity of the glycerolipid flux network, cultured embryos were incubated with [¹⁴C/¹³C]glycerol, and initial and steady state rates of [¹⁴C/¹³Cglyceryl] lipid accumulation were measured. At steady state, the linear accumulations of labeled lipid classes matched those expected from mass compositions. The system showed an apparently simple kinetic precursor–product relationship between the intermediate pool, dominated by diacylglycerol (DAG) and phosphatidylcholine (PC), and the triacylglycerol (TAG) product. We also conducted isotopomer analyses on hydrogenated lipid class species. [¹³C₃glyceryl] labeling of DAG and PC, together with estimates of endogenous [¹²C₃glyceryl] dilution, showed that each biosynthetically active lipid pool is ~30% of the total by moles. This validates the concept that lipid sub-pools can describe lipid biosynthetic networks. By tracking the kinetics of [¹³C₃glyceryl] and [¹³C₂acyl] labeling, we observed two distinct TAG synthesis components. The major TAG synthesis flux (~75%) was associated with >95% of the DAG/PC intermediate pool, with little glycerol being metabolized to fatty acids, and with little dilution from endogenous glycerol; a smaller flux exhibited converse characteristics. This kinetic heterogeneity was further explored using postlabeling embryo dissection and differential lipid extractions. The minor flux was tentatively localized to surface cells across the whole embryo. Such heterogeneity must be recognized in order to construct accurate gene expression patterns and metabolic networks describing lipid biosynthesis in developing embryos.

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Research Organization:: Michigan State Univ., East Lansing, MI (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER)

Grant/Contract Number:: SC0018269

OSTI ID:: 1830024

Alternate ID(s):: OSTI ID: 1904390

Journal Information:: Journal of Biological Chemistry, Journal Name: Journal of Biological Chemistry Vol. 298 Journal Issue: 1; ISSN 0021-9258

Publisher:: ElsevierCopyright Statement

Country of Publication:: United States

Language:: English

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59 BASIC BIOLOGICAL SCIENCES
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Title: Kinetic complexities of triacylglycerol accumulation in developing embryos from Camelina sativa provide evidence for multiple biosynthetic systems

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References (49)

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