Abstract
Studies evaluating the uptake of long-chain fatty acids in Caulobacter crescentus are consistent with a protein-mediated process. Using oleic acid (C18:1) as a substrate, fatty acid uptake was linear for up to 15 min. This process was saturable giving apparent Vmax and Km values of 374 pmol oleate transported/min/mg total protein and 61 μM oleate, respectively, consistent with the notion that one or more proteins are likely involved. The rates of fatty acid uptake in C. crescentus were comparable to those defined in Escherichia coli. Uncoupling the electron transport chain inhibited oleic acid uptake, indicating that like the long-chain fatty acid uptake systems defined in other gram-negative bacteria, this process is energy-dependent in C. crescentus. Long-chain acyl CoA synthetase activities were also evaluated to address whether vectorial acylation represented a likely mechanism driving fatty acid uptake in C. crescentus. These gram-negative bacteria have considerable long-chain acyl CoA synthetase activity (940 pmol oleoyl CoA formed/min/mg total protein), consistent with the notion that the formation of acyl CoA is coincident with uptake. These results suggest that long-chain fatty acid uptake in C. crescentus proceeds through a mechanism that is likely to involve one or more proteins.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Abumrad N, Harmon C, Ibrahimi A (1998) Membrane transport of long-chain fatty acids: evidence for a facilitated process. J Lipid Res 39:2309–2318
Azizan A, Black P (1994) Use of transposon TnphoA to identify genes for cell envelope proteins of Escherichia coli required for long-chain fatty acid transport: the periplasmic protein Tsp potentiates long-chain fatty acid transport. J Bacteriol 176:6653–6662
Azizan A, Sherin D, DiRusso C, Black P (1999) Energetics underlying the process of long-chain fatty acid transport. Arch Biochem Biophys 365:299–306
Bar-Tana J, Rose G, Shapiro B (1971) The purification and properties of microsomal Palmitoyl-coenzyme A synthetase. Biochem J 122:353–362
Black P, DiRusso C, Metzger A, Heimert T (1992) Cloning, sequencing, and expression of the fadD gene of Escherichia coli encoding Acyl coenzyme A synthetase. J Biol Chem 267:25513–25520
Black P, Zhang Q, Weimar J, DiRusso C (1997) Mutational analysis of a fatty acyl-coenzyme A synthetase signature motif identifies seven amino acid residues that modulate fatty acid substrate specificity. J Biol Chem 272:4896–4903
Black P, DiRusso C, Sherin D, MacColl R, Knudsen J, Weimar J (2000) Affinity labeling fatty acyl CoA synthetase with 9-p-azidophenoxy nonanoic acid and the identification of the fatty acid-binding site. J Biol Chem 275:38547–38553
DiRusso C, Black P (1999) Long chain fatty acid transport in bacteria and yeast. Paradigms for defining the mechanism underlying this protein-mediated process. Mol Cell Biochem 192:41–52
DiRusso C, Black P, Weimar J (1999) Molecular inroads into the regulation and metabolism of fatty acid; lessons from bacteria. Prog Lipid Res 38:129–197
Ginsburgh C, Black P, Nunn W (1984) Transport of long chain fatty acids in Escherichia coli. J Biol Chem 259:8437–8443
Hamilton J, Kamp F (1999) How are free fatty acids transported in membranes? Diabetes 48:2255–2269
Hottes A, Meewan M, Yang D, Arana N, Romero P, McAdams H, Stephens C (2004) Transcriptional profiling of Caulobacter crescentus during growth on complex and minimal media. J Bacteriol 186:1448–1461
Johnson R, Ely B (1977) Isolation of spontaneously derived mutants of Caulobacter crescentus. Genetics 86:25–32
Kumar G, Black P (1991) Linker mutagenesis of a bacterial fatty acid transport protein. J Biol Chem 266:1348–1353
Kumar G, Black P (1993) Bacterial long-chain fatty acid transport. Identification of amino acid residues within the outer membrane protein FadL required for activity. J Biol Chem 268:15469–15476
Lagenaur C, Agabian N (1978) Caulobacter flagellar organelle: synthesis, compartmentation, and assembly. J Bacteriol 135:1062–1069
Lawler M, Brun Y (2007) Advantages and mechanisms of polarity and cell shape determination in Caulobacter crescentus. Curr Opin Microbiol 10:630–637
Lowey B, Marczynski G, Dingwall A, Shapiro L (1990) Regulatory interactions between phospholipids synthesis and DNA replication in Caulobacter crescentus. J Bacteriol 172:5523–5530
Maloy S, Ginsburgh C, Simons R, Nunn W (1981) Transport of long and medium chain fatty acids by Escherichia coli K12. J Biol Chem 256:3735–3742
Mansour J, Henry S, Shapiro L (1980) Differential membrane phospholipid synthesis during the cell cycle of Caulobacter crescentus. J Bacteriol 143:262–269
Mansour J, Henry S, Shapiro L (1981) Phospholipid biosynthesis is required for stalk elongation in Caulobacter crescentus. J Bacteriol 145:1404–1409
Miller J (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY
Nierman W et al (2001) Complete genome sequence of Caulobacter crescentus. Proc Natl Acad Sci USA 98:4136–4141
O’Connell M, Henry S, Shapiro L (1986) Fatty acid degradation in Caulobacter crescentus. J Bacteriol 168:49–54
van den Berg B, Black P, Clemons W, Rapoport T (2004) Crystal structure of the long-chain fatty acid transporter FadL. Science 304:1506–1509
Acknowledgments
Special thanks go to Chris Petteys, who assisted on preliminary experiments, and to the members of the laboratory of Paul Black and Concetta DiRusso for helpful advice.
Author information
Authors and Affiliations
Corresponding author
Additional information
Communicated by Theo Hansen.
Rights and permissions
About this article
Cite this article
Zalatan, F., Black, P. Characterization of long-chain fatty acid uptake in Caulobacter crescentus . Arch Microbiol 193, 479–487 (2011). https://doi.org/10.1007/s00203-011-0694-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00203-011-0694-9