T-cell protein tyrosine phosphatase gives rise to two splice isoforms: TC48, which is localized to the endoplasmic reticulum (ER) and TC45, a nuclear protein. The present study was undertaken to identify proteins that are involved in targeting TC48 to the ER. We identified two TC48-interacting proteins, p25 and p23, from a yeast two-hybrid screen. p23 and p25 are members of a family of putative cargo receptors that are important for vesicular trafficking between Golgi complex and ER. Both p23 and p25 associate with overexpressed TC48 in Cos-1 cells as determined by coimmunoprecipitation. A significant amount of TC48 colocalized initially with ERGIC and Golgi complex markers (in addition to ER and nuclear membrane localization) and was then retrieved to the ER. Coexpression with p25 enhanced ER localization of TC48, whereas coexpression with p23 resulted in its trapping in membranous structures. Coexpression of a p25 mutant lacking the ER-localization signal KKxx resulted in enhanced Golgi localization of TC48. Forty C-terminal amino acid residues of TC48 (position 376-415) were sufficient for interaction with p23 (but not with p25) and targeted green fluorescence protein (GFP) to the Golgi complex. Targeting of GFP to the ER required 66 C-terminal amino acid residues of TC48 (position 350-415), which showed interaction with p25 and p23. We suggest that TC48 translocates to the Golgi complex along the secretory pathway, whereas its ER localization is maintained by selective retrieval enabled by interactions with p25 and p23.