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The Protein Journal

, 28:362 | Cite as

Hydrophobic Interactions Stabilize the Basigin-MCT1 Complex

  • NiCole A. Finch
  • Paul J. Linser
  • Judith D. OchrietorEmail author
Article

Abstract

Previous reports demonstrated that monocarboxylate transporter-1 (MCT1) interacts with Basigin. It was hypothesized that the two proteins interact via the transmembrane domain of Basigin, specifically through the glutamate residue within the domain. We therefore sought to test this hypothesis and determine which amino acids of the Basigin protein are necessary for the interaction with MCT1. Probes consisting of the full-length putative transmembrane domain, as well as small regions of the domain, were generated for use in ELISA binding assays using endogenous mouse MCT1. Site directed mutagenesis of candidate residues was performed and probes were generated for ELISA analyses to determine the specific residues involved. The data suggest that hydrophobic residues at the N- and C-termini of the putative transmembrane domain of Basigin interact with MCT1, but the glutamate plays no role. The previously proposed hypothesis is partially correct, in that the putative transmembrane domain of Basigin does interact with MCT1.

Keywords

Basigin Monocarboxylate transporter 1 Metabolism Retina ELISA 

Abbreviations

AP

Alkaline phosphatase

BSA

Bovine serum albumin

CD

Cluster of differentiation

cDNA

Complementary deoxyribonucleic acid

ELISA

Enzyme-linked immunosorbant assay

EMMPRIN

Extracellular matrix metalloproteinase inducer

FRET

Fluorescence resonance energy transfer

Ig

Immunoglobulin

IPTG

Isopropyl β-d-thiogalactopyranoside

MCT

Monocarboxylate transporter

PBS

Phosphate buffered saline

PCR

Polymerase chain reaction

RPE

Retinal pigmented epithelium

TM

Transmembrane domain

6XHis

Six C-terminal histidines

Notes

Acknowledgments

This work was supported by NIH F32EY13918 and UNF Academic Affairs (to JDO) and NSF IBN-0113697 (to PJL). We thank Dr. Andrea Kohn, University of Florida Whitney Laboratory for Marine Biosciences and Dr. Michael Lentz, University of North Florida for constructive criticisms. We also thank Mr. Greggory Wilhoite, Ms. Bahishta Yaqubi, and Mr. Anthony Vinson for technical assistance.

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Copyright information

© Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • NiCole A. Finch
    • 1
  • Paul J. Linser
    • 2
  • Judith D. Ochrietor
    • 1
    Email author
  1. 1.Department of BiologyUniversity of North FloridaJacksonvilleUSA
  2. 2.Whitney Laboratory for Marine Biosciences and the Department of Anatomy and Cell BiologyUniversity of FloridaSt. AugustineUSA

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