The ADP/ATP carrier is the 32-kilodalton receptor for an NH2-terminally myristylated src peptide but not for pp60src polypeptide.

Abstract

Membrane binding of pp60src is initiated via its myristylated NH2 terminus. To identify a candidate pp6G' docking protein or receptor in the membrane, a radiolabelled peptide corresponding to the pp6tO' NH2-terminal membrane binding domain was cross-linked to fibroblast membranes and found to specifically label a 32-kDa protein. This protein was purified by appending an affinity tag to the peptide probe so that the cross-linked complex could be isolated via affinity chromatography. Microsequencing indicated that the 32-kDa protein was the mitochondrial ADP/ATP carrier (AAC). This result was further confirmed by the ability of an antibody to the AAC to immunoprecipitate the cross-linked complex, by the ability of certain inhibitors of the AAC to block cross-linking, and by membrane fractionation to show that complex formation occurred essentially exclusively in the mitochondrial fraction. While the AAC bound the myristyl-src peptide in a specific manner both in vitro and in vivo, its localization to the inner membrane of the mitochondrion precludes its being a pp60" binding protein. An analysis of pp6OV' binding in vitro was consistent with this expectation. Thus, use of a myristyl-src peptide revealed an unexpected and previously unidentified binding capacity of the AAC, most likely related to the ability of long-chain fatty acyl coenzyme As to serve as AAC inhibitors. The amphipathic nature of the pp6@' NH2 terminus suggests alternative strategies for uncovering pp6G0 membrane binding species.Cellular transformation by the Rous sarcoma virus (RSV) is mediated by the membrane-bound tyrosine kinase pp6ov-srC, a

The Interaction between Cell Adhesion Molecule L1, Matrix Metalloproteinase 14, and Adenine Nucleotide Translocator at the Plasma Membrane Regulates L1-Mediated Neurite Outgrowth of Murine Cerebellar Neurons

“…Cell surface topology of ANT So far, only three studies on Leishmania, murine fibroblasts, and breast carcinoma and fibrosarcoma cells provide solid evidence that ANT is not only localized in mitochondria, but also at the cell surface (Sigal and Resh, 1993;Detke and Elsabrouty, 2008;Radichev et al, 2009). Our study confirms these findings and shows that the three ANT loops that project into the lumen of mitochondria are exposed at the neuronal cell surface (Fig.…”

“…Since it has been reported that ANT is located at the plasma membrane of Leishmania, fibroblasts, and cancer cells (Sigal and Resh, 1993;Detke and Elsabrouty, 2008;Radichev et al, 2009), and since we demonstrated that ANT interacts with the extracellular domain of L1, we tested whether ANT1 and/or ANT2 are present at the cell surface of cultured cerebellar neurons. Therefore, live primary cultures of cerebellar neurons were subjected to cell surface biotinylation, and biotinylated proteins were isolated using streptavidin-conjugated beads.…”

Permeabilization of the mitochondrial inner membrane during apoptosis: impact of the adenine nucleotide translocator

“…63 However, at present it is not clear whether Figure 1 Molecules influencing ANT-mediated pore formation in chemically defined systems. The upper part lists proteins affecting the pore function of ANT, whereas the lower part focuses on non-protein molecules.…”

Amino-terminal basic residues of Src mediate membrane binding through electrostatic interaction with acidic phospholipids.

“…Crosslinking experiments identified a 32-kDa protein as the major species labeled by a myristoylated amino-terminal Src peptide probe (12,13). However, upon purification, this protein was found to be the ADP/ATP carrier, a mitochondrial protein capable of binding myristoyl-Src peptide but not the Src protein (14). To date no other protein has been identified as a binding partner for the Src amino terminus.…”