Skip to main content
SpringerLink
Log in

Functional Purification of the Monocarboxylate Transporter of the Yeast Candida utilis

  • Original Paper
  • Published:
Biotechnology Letters Aims and scope Submit manuscript

Abstract

Plasma membranes of the yeast, Candida utilis, were solubilized with octyl-β-d-glucopyranoside and a fraction enriched in the lactate carrier was obtained with DEAE-Sepharose anion-exchange chromatography, after elution with 0.4 M NaCl. The uptake of lactic acid into proteoliposomes, containing the purified protein fraction and cytochrome c oxidase, was dependent on a proton-motive force and the transport specificity was consistent with the one of C. utilis intact cells. Overall, we have obtained a plasma membrane fraction enriched in the lactate carrier of C. utilis in which the transport properties were preserved. Given the similarities between the lactate transport of C. utilis and the one of mammalian cells, this purified system could be further explored to screen for specific lactate inhibitors, with potential therapeutic applications.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Allen PJ, Brooks GA (1994) Partial purification and reconstitution of the sarcolemmal l-lactate carrier from rat skeletal muscle. Biochem J 303:207–212

    PubMed  CAS  Google Scholar 

  • Casal M, Leão C (1995) Utilization of short-chain monocarboxylic acids by the yeast Torulaspora delbrueckii: specificity of the transport systems and their regulation. Biochim Biophys Acta 1267:122–130

    Article  PubMed  Google Scholar 

  • Casal M, Paiva S, Andrade RP, Gancedo C, Leão C (1999) The lactate-proton symport of Saccharomyces cerevisiae is encoded by JEN1. J Bacteriol 181:2620–2623

    PubMed  CAS  Google Scholar 

  • Cássio F, Leão C (1991) Low- and high-affinity transport systems for citric acid in the yeast Candida utilis. Appl Environ Microbiol 57:3623–3628

    PubMed  Google Scholar 

  • Cássio F, Leão C (1993) A comparative study on the transport of l(−)malic acid and other short-chain carboxylic acids in the yeast Candida utilis: evidence for a general organic acid permease. Yeast 9:743–752

    Article  PubMed  Google Scholar 

  • Cássio F, Corte-Real M, Leão C (1993) Quantitative analysis of proton movements associated with the uptake of weak carboxylic acids. The yeast Candida utilis as a model. Biochim Biophys Acta 1153:59–66

    Article  PubMed  Google Scholar 

  • Cássio F, Leão C, van Uden N (1987) Transport of lactate and other short-chain monocarboxylates in the yeast Saccharomyces cerevisiae. Appl Environ Microbiol 53:509–513

    PubMed  Google Scholar 

  • Gerós H, Cássio F, Leão C (1996) Reconstitution of lactate proton symport activity in plasma membrane vesicles from the yeast Candida utilis. Yeast 12:1263–1272

    Article  PubMed  Google Scholar 

  • Gerós H, Cássio F, Leão C (2000a) Utilization and transport of acetic acid in Dekkera anomala and their implications on the survival of the yeast in acidic environments. J Food Prot 63:96–101

    Google Scholar 

  • Gerós H, Baltazar F, Cássio F, Leão C (2000b) l-[U-14C]Lactate binding to a 43 kDa protein in plasma membranes of Candida utilis. Microbiology 146:695–699

    Google Scholar 

  • Halestrap AP, Meredith D (2004) The SLC16 gene family-from monocarboxylate transporters (MCTs) to aromatic amino acid transporters and beyond. Pflugers Arch 447:619–628

    Article  PubMed  CAS  Google Scholar 

  • Jennings ML, Adams-Lackey M (1982) A rabbit erythrocyte membrane protein associated with l-lactate transport. J Biol Chem 257:12866–12871

    PubMed  CAS  Google Scholar 

  • Kirk P, Wilson MC, Heddle C, Brown MH, Barclay AN, Halestrap AP (2000) CD147 is tightly associated with lactate transporters MCT1 and MCT4 and facilitates their cell surface expression. EMBO J 19:3896–3904

    Article  PubMed  CAS  Google Scholar 

  • Leão C, van Uden N (1986) Transport of lactate and other short-chain monocarboxylates in the yeast Candida utilis. Appl Microbiol Biotechnol 23:389–393

    Article  Google Scholar 

  • Makuc J, Paiva S, Schauen M, Krämer R, André B, Casal M, Leão C, Boles E (2001) The putative monocarboxylate permeases of the yeast Saccharomyces cerevisiae do not transport monocarboxylic acids across the plasma membrane. Yeast 18:1131–1143

    Article  PubMed  CAS  Google Scholar 

  • Matsushita K, Patel L, Gennis RB, Kaback HR (1983) Reconstitution of active transport in proteoliposomes containing cytochrome c oxidase and lac carrier protein purified from Escherichia coli. Proc Natl Acad Sci 80:4889–4893

    Article  PubMed  CAS  Google Scholar 

  • McCullagh KJA, Bonen A (1995) l(+)-Lactate binding to a protein in rat skeletal muscle plasma membranes. Can J Appl Physiol 20:112–124

    PubMed  CAS  Google Scholar 

  • Poole RC, Halestrap AP (1992) Identification and partial purification of the erythrocyte l-lactate transporter. Biochem J 283:855–862

    PubMed  CAS  Google Scholar 

  • Poole RC, Halestrap AP (1994) N-terminal sequence analysis of the rabbit erythrocyte lactate transporter suggests identity with the cloned monocarboxylate transporter protein MCT1. Biochem J 303:755–759

    PubMed  CAS  Google Scholar 

  • Soares-Silva I, Schuller D, Andrade RP, Baltazar F, Cássio F, Casal M (2003) Functional expression of the lactate permease Jen1p of Saccharomyces cerevisiae in Pichia pastoris. Biochem J 376:781–787

    Article  PubMed  CAS  Google Scholar 

  • Van Leeuwen CCM, Postma E, Van der Broek PJA, Steveninck J (1991) Proton-motive force-driven d-galactose transport in plasma membrane vesicles from the yeast Kluyveromyces marxianus. J Biol Chem 266:12146–12151

    PubMed  Google Scholar 

  • Viitanen P, Newman NJ, Foster DL, Wilson TH, Kaback HR (1986) Purification, reconstitution and characterization of the lac permease of Escherichia coli. Method Enzymol 125:429–452

    Article  CAS  Google Scholar 

  • Welch SG, Metcalfe HK, Monson JP, Cohen RD, Henderson RM, Iles RA (1984) l(+)-Lactate binding to preparations of rat hepatocyte plasma membranes. J␣Biol Chem 259:15264–15271

    PubMed  CAS  Google Scholar 

Download references

Acknowledgement

F. B. received a post-doctoral fellowship from Fundação para a Ciência e a Tecnologia (SFRH/BPD/7170/2001).

Author information

Authors and Affiliations

  1. Life and Health Sciences Research Institute (ICVS), School of Health Sciences, University of Minho, Braga, Portugal

    Fátima Baltazar & Cecília Leão

  2. Department of Biology, Biology Centre, University of Minho, Braga, Portugal

    Fernanda Cássio

Authors
  1. Fátima Baltazar
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Fernanda Cássio
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Cecília Leão
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Fátima Baltazar.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Baltazar, F., Cássio, F. & Leão, C. Functional Purification of the Monocarboxylate Transporter of the Yeast Candida utilis . Biotechnol Lett 28, 1221–1226 (2006). https://doi.org/10.1007/s10529-006-9088-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10529-006-9088-5

Keywords

Search

Navigation