Functional and Physical Interaction between Human Lactate Dehydrogenase B and Na+/H+ Exchanger Isoform 1

Abstract

The ubiquitous plasma membrane Na+/H+ exchanger 1 (NHE1) is rapidly activated in response to various extracellular stimuli and maintains normal cytoplasmic pH. Yeast two‐hybrid screening was used in order to identify proteins interacting with NHE1 using its cytoplasmic domain as a bait from HeLa cDNA library. One of the interacting cDNA clones was human Lactate dehydrogenase B (LDHB). In vitro translated LDHB was pulled down together with GST‐NHE1.cd protein in the GST pull down assay, confirming the interaction in vitro. LDHB antibody immuno‐precipitated endogenous LDHB together with NHE1 from H9c2 cells, validating cellular interaction between NHE1 and LDHB. Subsequent analysis revealed that the overexpression of LDHB increased intracellular PH, implying opening of the NHE1 transporter. Moreover, overexpression of LDHB activated caspase 3 and induced cell death, consistent with the expected phenotype of hyper‐activation of NHE1. Collectively, our data indicate that LDHB modulates NHE1 activity via physical interaction.