Purification and characterization of pituitary bovine somatotropin

D. C. Wood; W. J. Salsgiver; T. R. Kasser; G. W. Lange; E. Rowold; B. N. Violand; A. Johnson; R. M. Leimgruber; G. R. Parr; N. R. Siegel; N. M. Kimack; C. E. Smith; J. F. Zobel; M. Ganguli; J. R. Garbow; G. Bild; G. G. Krivi

Purification and characterization of pituitary bovine somatotropin

D. C. Wood
, W. J. Salsgiver
, T. R. Kasser
, G. W. Lange
, E. Rowold
, B. N. Violand
, A. Johnson
, R. M. Leimgruber
, G. R. Parr
, N. R. Siegel
, N. M. Kimack
, C. E. Smith
, J. F. Zobel
, M. Ganguli
, J. R. Garbow
, G. Bild
, G. G. Krivi

Research output: Contribution to journal › Article › peer-review

Abstract

Bovine somatotropin (bST) has been isolated from pituitary glands and compared in a variety of chemical analyses and bioassays with somatotropin derived from recombinant Escherichia coli. Comparison of pituitary extracts and purified bST by Western blot analysis of two-dimensional gels suggested that the immunoreactive somatotropin species present in the extract were also present in the purified material, with no significant losses or degradation as a result of the purification method. NH₂-terminal sequence analysis indicated the presence of equal quantities of Ala-Phe-Pro-Ala-Met-Ser-Leu-Ser- and Phe-Pro-Ala-Met-Ser-Leu-Ser- sequences. The Met-Ser-Leu-Ser-NH₂-terminal sequence, a degradation product observed in NIH standard lots, was not detected. Assay of bioactivity in a bovine liver receptor-binding assay and in a female rate growth assay showed pituitary bST and recombinant methionyl-bovine somatotropin to be equipotent. Tryptic maps and sequence analysis of pituitary-derived somatotropin suggest the presence of isoaspartate derivatization at Asp¹²⁸.

Original language	English
Pages (from-to)	14741-14747
Number of pages	7
Journal	Journal of Biological Chemistry
Volume	264
Issue number	25
State	Published - 1989

Cite this

Wood, D. C., Salsgiver, W. J., Kasser, T. R., Lange, G. W., Rowold, E., Violand, B. N., Johnson, A., Leimgruber, R. M., Parr, G. R., Siegel, N. R., Kimack, N. M., Smith, C. E., Zobel, J. F., Ganguli, M., Garbow, J. R., Bild, G., & Krivi, G. G. (1989). Purification and characterization of pituitary bovine somatotropin. Journal of Biological Chemistry, 264(25), 14741-14747.

@article{9696eb0ef9ee4644a1c61b6888ada1a3,

title = "Purification and characterization of pituitary bovine somatotropin",

abstract = "Bovine somatotropin (bST) has been isolated from pituitary glands and compared in a variety of chemical analyses and bioassays with somatotropin derived from recombinant Escherichia coli. Comparison of pituitary extracts and purified bST by Western blot analysis of two-dimensional gels suggested that the immunoreactive somatotropin species present in the extract were also present in the purified material, with no significant losses or degradation as a result of the purification method. NH2-terminal sequence analysis indicated the presence of equal quantities of Ala-Phe-Pro-Ala-Met-Ser-Leu-Ser- and Phe-Pro-Ala-Met-Ser-Leu-Ser- sequences. The Met-Ser-Leu-Ser-NH2-terminal sequence, a degradation product observed in NIH standard lots, was not detected. Assay of bioactivity in a bovine liver receptor-binding assay and in a female rate growth assay showed pituitary bST and recombinant methionyl-bovine somatotropin to be equipotent. Tryptic maps and sequence analysis of pituitary-derived somatotropin suggest the presence of isoaspartate derivatization at Asp128.",

author = "Wood, \{D. C.\} and Salsgiver, \{W. J.\} and Kasser, \{T. R.\} and Lange, \{G. W.\} and E. Rowold and Violand, \{B. N.\} and A. Johnson and Leimgruber, \{R. M.\} and Parr, \{G. R.\} and Siegel, \{N. R.\} and Kimack, \{N. M.\} and Smith, \{C. E.\} and Zobel, \{J. F.\} and M. Ganguli and Garbow, \{J. R.\} and G. Bild and Krivi, \{G. G.\}",

year = "1989",

language = "English",

volume = "264",

pages = "14741--14747",

journal = "Journal of Biological Chemistry",

issn = "0021-9258",

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TY - JOUR

T1 - Purification and characterization of pituitary bovine somatotropin

AU - Wood, D. C.

AU - Salsgiver, W. J.

AU - Kasser, T. R.

AU - Lange, G. W.

AU - Rowold, E.

AU - Violand, B. N.

AU - Johnson, A.

AU - Leimgruber, R. M.

AU - Parr, G. R.

AU - Siegel, N. R.

AU - Kimack, N. M.

AU - Smith, C. E.

AU - Zobel, J. F.

AU - Ganguli, M.

AU - Garbow, J. R.

AU - Bild, G.

AU - Krivi, G. G.

PY - 1989

Y1 - 1989

N2 - Bovine somatotropin (bST) has been isolated from pituitary glands and compared in a variety of chemical analyses and bioassays with somatotropin derived from recombinant Escherichia coli. Comparison of pituitary extracts and purified bST by Western blot analysis of two-dimensional gels suggested that the immunoreactive somatotropin species present in the extract were also present in the purified material, with no significant losses or degradation as a result of the purification method. NH2-terminal sequence analysis indicated the presence of equal quantities of Ala-Phe-Pro-Ala-Met-Ser-Leu-Ser- and Phe-Pro-Ala-Met-Ser-Leu-Ser- sequences. The Met-Ser-Leu-Ser-NH2-terminal sequence, a degradation product observed in NIH standard lots, was not detected. Assay of bioactivity in a bovine liver receptor-binding assay and in a female rate growth assay showed pituitary bST and recombinant methionyl-bovine somatotropin to be equipotent. Tryptic maps and sequence analysis of pituitary-derived somatotropin suggest the presence of isoaspartate derivatization at Asp128.

AB - Bovine somatotropin (bST) has been isolated from pituitary glands and compared in a variety of chemical analyses and bioassays with somatotropin derived from recombinant Escherichia coli. Comparison of pituitary extracts and purified bST by Western blot analysis of two-dimensional gels suggested that the immunoreactive somatotropin species present in the extract were also present in the purified material, with no significant losses or degradation as a result of the purification method. NH2-terminal sequence analysis indicated the presence of equal quantities of Ala-Phe-Pro-Ala-Met-Ser-Leu-Ser- and Phe-Pro-Ala-Met-Ser-Leu-Ser- sequences. The Met-Ser-Leu-Ser-NH2-terminal sequence, a degradation product observed in NIH standard lots, was not detected. Assay of bioactivity in a bovine liver receptor-binding assay and in a female rate growth assay showed pituitary bST and recombinant methionyl-bovine somatotropin to be equipotent. Tryptic maps and sequence analysis of pituitary-derived somatotropin suggest the presence of isoaspartate derivatization at Asp128.

UR - https://www.scopus.com/pages/publications/0024457384

M3 - Article

C2 - 2768239

AN - SCOPUS:0024457384

SN - 0021-9258

VL - 264

SP - 14741

EP - 14747

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

IS - 25

ER -

Purification and characterization of pituitary bovine somatotropin

Abstract

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