Induction of novel Grp75 isoforms by 2-deoxyglucose in human and murine fibroblasts

Grp75 is a stress-inducible mitochondrial chaperone which has a high homology to senescence-related protein, p66mot mortalin. In human cells the mortalin gene assigns to the locus of a putative tumor suppressor gene for myeloid malignancies. In order to study expression and localization of Grp75 and p66mot in human and murine fibroblast lines, polyclonal antibodies were raised to conserved portions of each sequence. HT1080 and C3H10T1/2 cells were treated with various Grp-inducing agents. A single 75 kDa band was detected by Western blot of cytoplasmic proteins which was not greatly altered after thermal stress or treatment with l-azetidine-2-carboxylic acid or nonactin. However, glucose deprivation by 2-deoxyglucose treatment induced five novel isoforms at 74–75 kDa mass. Mortalin at 66 kDa could not be detected under these treatment conditions.