Abstract
Pancreatic thread protein (PTP) forms double helical threads in the neutralpH range after purification, undergoing freely reversible,pH-dependent globule-fibril transformation. The purified bovine PTP consists on SDS gels of two carbohydrate-free polypeptide chains (Grosset al., 1985). Plasma desorption mass spectrometry and amino acid sequence analysis now confirm that bovine PTP contains two disulfide-bonded polypeptides, an A chain of 101 amino acid residues with a molecular weight of 11,073 and a B chain of 35 residues with a molecular weight of 3970. The intact protein exhibits a molecular weight of 15,036, agreeing >99.9% with the molecular weight calculated from the sequence. The B chain sequence was determined by gas-phase Edman degradation of the intact polypeptide. The A chain sequence was determined from overlapping peptides generated by cleavage at lysyl, tryptophanyl, and aspartyl-prolyl residues. Based upon the bovine PTP cDNA structure, the two chains of the protein result from cleavage of a single polypeptide with removal of a dipeptide between the NH2-terminal A chain and COOH-terminal B chain. Comparison of bovine PTP with other proteins reveals significant structural relatedness with the single-chain homologues from human and rat pancreas and with the motif associated with Ca2+-dependent carbohydrate recognition domains. The physiological role of PTP has not yet been resolved. The protein is present in very high concentration in pancreatic secretion and it has been detected in brain lesions in Alzheimer's disease and Down syndrome and in regenerating rat pancreatic islets. The present results provide a firm protein base for ongoing molecular, physical-chemical, and structure-function studies of this unusual protein.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Crabb, J. W., Armes, L. G., Carr, S. A., Johnson, C. M., Roberts, G. D., Bordoli, R. S., and McKeehan, W. L. (1986).Biochemistry 25, 4988–4993.
Crabb, J. W., Johnson, C. M., Carr, S. A., Armes, L. G., and Saari, J. C. (1988).J. Biol. Chem. 263, 18,678–18,687.
Chou, J. Y., and Fasman, G. D. (1978).Adv. Enzymol. 47, 145–148.
De Caro, A. M., Bonicel, J. J., Rouimi, P., De Caro, J. D., Sarles, H., and Rovery, M. (1987).Eur. J. Biochem. 168, 201–207.
Drickamer, K. (1988).J. Biol. Chem. 263, 9557–9560.
Fairbanks, G., Steck, T. L., and Wallach, F. H. (1971).Biochemistry 10, 2606–2617.
Fontana, A. (1972).Methods Enzymol. 25, 419–423.
Friedman, M., Krull, L. H., and Cavins, J. F. (1970).J. Biol. Chem. 245, 3868–3871.
Fullmer, C. S. (1984).Anal. Biochem. 142, 336–339.
Gross, J. (1949).J. Exp. Med. 89, 699–708.
Gross, J. (1952).Fed. Proc. 11, 60 (abstract).
Gross, J. (1951).Proc. Soc. Exp. Biol. Med. 78, 241–244.
Gross, J., Brauer, A. W., Bringhurst, R. F., Corbett, C., and Margolis, M. N. (1985a).Proc. Natl. Acad. Sci. USA 82, 5627–5631.
Gross, J., Carlson, R. J., Brauer, A. W., Margolis, M. N., Warshaw, A. W., and Wands, J. R. (1985b).J. Clin. Invest. 76, 2115–2126.
Hampton, B., Burgess, W. H., Marshak, D. R. Cullen, K. J., and Perdue, J. F. (1989).J. Biol. Chem. 264, 19,155–19,160.
Hopp, T. P., and Woods, K. R. (1981).Proc. Natl. Acad. Sci. USA 78, 3824–3828.
Laemmli, U. K. (1970).Nature 227, 680–685.
Ozturk, M., De LaMonte, S. M., Gross, J., and Wands, J. R. (1989).Proc. Natl. Acad. Sci. USA 68, 419–423.
Patthy, L. (1988).Biochem. J. 253, 309–311.
Pearson, W. R., and Lipman, D. J. (1988).Proc. Natl. Acad. Sci. USA 85, 2444–2448.
Petersen, T. E. (1988).FEBS Lett. 231, 51–53.
Sundqvist, B., Roepstorff, P., Fohlman, J., Hedin, A., Häkansson, P., Kamensky, I., Lingberg, M., Salehpour, M., and Säwe, G. (1984).Science 226, 696–698.
Swank, R. T., and Munkres, K. D. (1971).Anal. Biochem. 39, 462–477.
Tanzi, R. E., Gusella, J. F., Watkins, P. C., Bruns, G. A. P., St. George-Hyslop, P., Van Keuren, M. L., Patterson, D., Pagan, S., Kurnit, D. M., and Neve, R. L. (1987).Science 235, 880–884.
Tarr, G. E. (1986). InMicrocharacterization of Polypeptides, A Practical Manual (Shively, J. E., ed.), Humana Press, Clifton, New Jersey, pp. 155–194.
Terazono, K., Yamamoto, H., Takasawa, S., Shiga, K., Yonemura, Y., Tochino, Y., and Okamoto, H. (1988).J. Biol. Chem. 263, 2111–2114.
West, K. A., and Crabb, J. W. (1990). InCurrent Research in Protein Chemistry (Villafranca, J. J., ed.), Academic Press, San Diego, pp. 37–48.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Cai, L., Harris, W.R., Marshak, D.R. et al. Structural analysis of bovine pancreatic thread protein. J Protein Chem 9, 623–632 (1990). https://doi.org/10.1007/BF01025016
Received:
Published:
Issue Date:
DOI: https://doi.org/10.1007/BF01025016