Purification and structure of porcine calcitonin-1

Abstract

We wish to report the isolation in pure form of a porcine calcitonin (PC-1) and the elucidation of its structure.1(1) We also wish to acknowledge the valuable assistance of C. Pidacks for chromatography development work, as well as E. Lindemann and H. Falk for biological assay data.

Isolation and Determination of the Amino Acid Sequence of Chicken Calcitonin I from Chicken Ultimobranchial Glands

“…The subsequence [1][2][3][4][5][6][7][8][9][10][11][12][13][14] of performic acid-oxidized chicken calcitonin I and the sequences of T-l, T-2, and T-3 were determined as shown in Table HE. T-4 was found to be the N-terminal peptide, since the amino acid composition of T-4 (Table II) was identical with that of subsequence [1][2][3][4][5][6][7][8][9][10][11] of performic acidoxidized chicken calcitonin I. Two half-cystine residues at the 1st and 7th positions form a disulfide linkage in the chicken calcitonin I, since the approximate molecular weight of chicken calcitonin I was 3,800 ( Fig.…”

“…7). As shown in Tables II and m, P-l contained the amino acids of subsequence [16][17][18][19][20][21][22][23][24][25][26][27][28][29][30][31][32] and P-2 contained the amino acids of subsequence [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15] of TABLE in. Sequence determination of chicken calcitonin I.…”

Characterization of the Immunochemical Forms of Calcitonin Released by a Medullary Thyroid Carcinoma in Tissue Culture

“…It could be demonstrated that a major proportion of the immunochemical enlargement is dependent upon intermolecular disulfide bridge formation whereas aggregation or non-convalent protein binding account for a smaller component of the heterogeneity. In view of the absence of binding of the immunoreactive material to the lectin agarose, carbohydrate side chains, at least of INTRODUCTION Calcitonin in several species is known to be a 32-amino acid polypeptide hormone (1)(2)(3)(4)(5)(6) containing an intrachain disulfide bridge between sequence positions 1 and 7 at the NH2-terminal end of the molecule and an amide group on the COOH-terminal proline. The human hormone (hCT)' is secreted by normal (7)(8)(9)(10)(11)(12) and neoplastic C cells (7)(8)(9)(10)(11)(12)(13)(14)(15), as well as by non-C cell neoplasms (16)(17)(18).…”

Calcitonin Activity of Particulate Fractions of the Ovine Thyroid Gland

“…It was determined through the use of sequential degradation by the phenylisothiocyanate procedure, digestion with exopeptidases, and isolation and analysis of peptide subfragments resulting from cleavage of the polypeptide by trypsin, pepsin, chymotrypsin, papain, cyanogen bromide, and limited hydrolysis in dilute acid and concentrated acid. Neher et al (1968) simultaneously arrived at the identical amino acid sequence for porcine CT and Bell et al (1968) arrived at the same sequence finding the weight-average molecular weight of porcine CT to be 3700, which was in excellent agreement with the value of 3604 as required by the amino acid sequence of the molecule. Distinctive structural features of the 32 amino acid molecule are that the proportion of charged amino acids is low, the carboxyl-terminal amino acid is prolinamide, and ~here is a 1-7 intrachain disulfide bridge, providing a 2J-membered ring at the amino terminus.…”