Figure 1.

Purification of bovine TPST (A) and structure of the human TPST-2 gene and mRNA (B). (A) A mixture of TPST isolated on a preparative scale from bovine adrenal medulla by affinity chromatography on glutathione S-transferase-sulfophilin and ¹²⁵I-labeled TPST purified on an analytical scale as described (17) was subjected to SDS/PAGE followed by autoradiography of the wet gel. The bracket indicates ¹²⁵I-labeled TPST. The corresponding region of the gel was excised and subjected to tryptic digestion followed by HPLC separation of the peptides and microsequencing. (B) Boxes above the TPST-2 mRNA show the putative exons of the TPST-2 gene in the BAC 445C9; numbers refer to the nucleotides of the BAC 445C9 (Z95115) corresponding to the 5′ and 3′ ends of the exons. The shaded area of the TPST-2 mRNA indicates the ORF. The lines below the TPST-2 mRNA indicate some of the TPST-2 EST clones in the database. Their accession numbers are given above the lines; numbers below the lines give the 5′ and 3′ ends of the EST sequences and refer to the nucleotide sequence of the TPST-2 mRNA. The asterisks and bold accession numbers indicate the exon and EST sequences, respectively, that encode the sequence of the tryptic peptide isolated from purified bovine adrenal medullary TPST.