title: Efficient targeting to storage granules of human proinsulins with altered propeptide domain.
abstract: In neuronal and endocrine cells, peptide hormones are selectively segregated into storage granules, while other proteins are exported continuously without storage. Sorting of hormones by cellular machinery involves the recognition of specific structural domains on prohormone molecules. Since the propeptide of insulin is known to play an important role in its three-dimensional structure, it is reasonable to speculate that targeting of proinsulin to storage granules would require a functional connecting peptide. To test this hypothesis, we constructed two mutations in human proinsulin with different predicted structures. In one mutation, Ins delta C, the entire C peptide was deleted, resulting in an altered insulin in which the B and the A chains are joined contiguously. In the other mutation, Ins/IGF, the C peptide of proinsulin was replaced with the unrelated 12-amino acid connecting peptide of human insulin-like growth factor-I; this substitution should permit correct folding of the B and A chains to form a tertiary structure similar to that of proinsulin. By several biochemical and morphological criteria, we found that Ins/IGF is efficiently targeted to storage granules, suggesting that the C peptide of proinsulin does not contain necessary sorting information. Unexpectedly, Ins delta C, which presumably cannot fold properly, is also targeted to granules at a high efficiency. These results imply that either the targeting machinery can tolerate changes in the tertiary structure of transported proteins, or that the B and A chains of insulin can form a relatively intact three-dimensional structure even in the absence of C peptide.
authors.affiliation: Department of Physiology-Anatomy, University of California, Berkeley 94720.
authors.forename: S K
authors.initials: SK
authors.lastname: Powell
authors.forename: L
authors.initials: L
authors.lastname: Orci
authors.forename: C S
authors.initials: CS
authors.lastname: Craik
authors.forename: H P
authors.initials: HP
authors.lastname: Moore
mesh_terms: D002451:Cell Compartmentation; D000242:Cyclic AMP / Q000502:physiology; D003594:Cytoplasmic Granules / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006801:Humans; D007150:Immunohistochemistry; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D007334:Insulin-Like Growth Factor I / Q000378:metabolism; D008970:Molecular Weight; D011384:Proinsulin / Q000378:metabolism*; D013329:Structure-Activity Relationship
chemical_list: D007328:Insulin; D007334:Insulin-Like Growth Factor I; D011384:Proinsulin; D000242:Cyclic AMP