PURIFICATION OF GOLGI ADAPTER PROTEIN-1 FROM BOVINE ADRENAL-GLAND AMDCHARACTERIZATION OF ITS BETA-1 (BETA') SUBUNIT BY MICROSEQUENCING

A method for the purification of the Golgi adaptor protein 1 from bovi ne adrenal gland tissue was devised to investigate the relationship of its beta 1 (formerly referred to as beta') subunit to known beta-type sequences. Adrenal gland tissue was chosen for this study because it yielded 2-3 times more adaptor protein 1 than a comparable preparation from bovine brain. Like its neuronal isoform, the beta 1 subunit from adrenal gland adaptor protein 1 is readily cleaved by trypsin into a 63-kDa N-terminal fragment and a 40-kDa C-terminal fragment, while the gamma subunit is largely refractory to digestion. Based on microseque ncing of 167 residues from the 63-kDa fragment, we noted 11 difference s to the corresponding region of the beta 2 (formerly beta) subunit of the plasma membrane adaptor protein 2, but only one difference to the corresponding region of a beta-type protein encoded by the rat cDNA c lone AP105a which is supposed to be a variant of the beta 2 subunit of the plasma membrane adaptor protein 2 [Kirchhausen, T, Nathanson, K. L., Matsui, W., Vaisberg, A., Chow, E. P., Burne, C., Keen, J. H. and Davis, A. E. (1989) Proc. Natl Acad. Sci. USA 84, 8805-8809]. Alignmen t of 187 residues from the 40-kDa beta 1 C-terminal fragment revealed differences in 77 positions to the corresponding region of the beta 2 subunit and differences in 23 positions compared to the supposed beta 2-like protein. These findings suggest that the protein encoded by the rat cDNA clone AP105a is more closely related to the beta 1 subunit o f the bovine adrenal Golgi adaptor protein 1 than to the beta 2 subuni t of the rat plasma membrane adaptor protein 2.