Identification of circulating growth hormone-binding proteins in domestic poultry

An initial characterization

Regina Vasilatos-Younken, B. J. Andersen, R. W. Rosebrough, J. P. McMurtry, W. L. Bacon

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Multiple growth hormone (GH)-binding proteins (GHBPs) were identified in serum and plasma samples from domestic chickens and turkeys. Proteins were separated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis on 10% acrylamide, 2.7% bis discontinuous gels under reducing conditions and electrotransferred to nitrocellulose paper. Western blots were incubated with 125I-labelled recombinant chicken GH (cGH) or bovine GH and GHBPs visualized by means of autoradiography. In fresh samples (<2 h from collection to gel electrophoresis), multiple minor high M(r) bands were evident between approximately 72000 and 175000. Two major bands were observed at approximately 69500 and 27500. The latter is consistent with previous reports for the rat and mouse serum GHBPs based on nucleotide sequence analysis. The minor bands were essentially undetectable after storage at -25°C for several months, and an additional major band at M approximately 52500 appeared. The M(r) 69500 major protein contained N-linked carbohydrate, as determined by a reduction in molecular size by treatment with peptide N-glycosidase F. Binding of 125I-labelled GH was partially inhibited by co-incubation with 50 μg unlabelled pituitary-derived cGH/ml and excess unlabelled porcine GH as well as ovine prolactin, but not by bovine insulin. Non-specific binding of 125I-labelled GH by serum albumin was also observed. A comparison was made between these GHBPs and the hepatic GH receptor (e.g. molecular weight estimates, affinity for homologous versus heterologous GHs, cross-reactivity with prolactin, presence of N-linked carbohydrate). The origin and relationship among the various molecular weight species of GHBPs identified, and their potential role in regulation of the biological activity of GH in birds, remain to be determined.

Original languageEnglish (US)
Pages (from-to)115-122
Number of pages8
JournalJournal of Endocrinology
Volume130
Issue number1
StatePublished - Jan 1 1991

Fingerprint

Poultry
Growth Hormone
Chickens
Prolactin
Molecular Weight
Gels
Carbohydrates
Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase
Somatotropin Receptors
Collodion
Acrylamide
Autoradiography
Serum
Serum Albumin
Sodium Dodecyl Sulfate
Birds
Sequence Analysis
Electrophoresis
somatotropin-binding protein
Polyacrylamide Gel Electrophoresis

All Science Journal Classification (ASJC) codes

  • Endocrinology, Diabetes and Metabolism
  • Endocrinology

Cite this

Vasilatos-Younken, Regina ; Andersen, B. J. ; Rosebrough, R. W. ; McMurtry, J. P. ; Bacon, W. L. / Identification of circulating growth hormone-binding proteins in domestic poultry : An initial characterization. In: Journal of Endocrinology. 1991 ; Vol. 130, No. 1. pp. 115-122.
@article{9b0d996a5f814f95b539f4755bf52cea,
title = "Identification of circulating growth hormone-binding proteins in domestic poultry: An initial characterization",
abstract = "Multiple growth hormone (GH)-binding proteins (GHBPs) were identified in serum and plasma samples from domestic chickens and turkeys. Proteins were separated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis on 10{\%} acrylamide, 2.7{\%} bis discontinuous gels under reducing conditions and electrotransferred to nitrocellulose paper. Western blots were incubated with 125I-labelled recombinant chicken GH (cGH) or bovine GH and GHBPs visualized by means of autoradiography. In fresh samples (<2 h from collection to gel electrophoresis), multiple minor high M(r) bands were evident between approximately 72000 and 175000. Two major bands were observed at approximately 69500 and 27500. The latter is consistent with previous reports for the rat and mouse serum GHBPs based on nucleotide sequence analysis. The minor bands were essentially undetectable after storage at -25°C for several months, and an additional major band at M approximately 52500 appeared. The M(r) 69500 major protein contained N-linked carbohydrate, as determined by a reduction in molecular size by treatment with peptide N-glycosidase F. Binding of 125I-labelled GH was partially inhibited by co-incubation with 50 μg unlabelled pituitary-derived cGH/ml and excess unlabelled porcine GH as well as ovine prolactin, but not by bovine insulin. Non-specific binding of 125I-labelled GH by serum albumin was also observed. A comparison was made between these GHBPs and the hepatic GH receptor (e.g. molecular weight estimates, affinity for homologous versus heterologous GHs, cross-reactivity with prolactin, presence of N-linked carbohydrate). The origin and relationship among the various molecular weight species of GHBPs identified, and their potential role in regulation of the biological activity of GH in birds, remain to be determined.",
author = "Regina Vasilatos-Younken and Andersen, {B. J.} and Rosebrough, {R. W.} and McMurtry, {J. P.} and Bacon, {W. L.}",
year = "1991",
month = "1",
day = "1",
language = "English (US)",
volume = "130",
pages = "115--122",
journal = "Journal of Endocrinology",
issn = "0022-0795",
publisher = "Society for Endocrinology",
number = "1",

}

Identification of circulating growth hormone-binding proteins in domestic poultry : An initial characterization. / Vasilatos-Younken, Regina; Andersen, B. J.; Rosebrough, R. W.; McMurtry, J. P.; Bacon, W. L.

In: Journal of Endocrinology, Vol. 130, No. 1, 01.01.1991, p. 115-122.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Identification of circulating growth hormone-binding proteins in domestic poultry

T2 - An initial characterization

AU - Vasilatos-Younken, Regina

AU - Andersen, B. J.

AU - Rosebrough, R. W.

AU - McMurtry, J. P.

AU - Bacon, W. L.

PY - 1991/1/1

Y1 - 1991/1/1

N2 - Multiple growth hormone (GH)-binding proteins (GHBPs) were identified in serum and plasma samples from domestic chickens and turkeys. Proteins were separated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis on 10% acrylamide, 2.7% bis discontinuous gels under reducing conditions and electrotransferred to nitrocellulose paper. Western blots were incubated with 125I-labelled recombinant chicken GH (cGH) or bovine GH and GHBPs visualized by means of autoradiography. In fresh samples (<2 h from collection to gel electrophoresis), multiple minor high M(r) bands were evident between approximately 72000 and 175000. Two major bands were observed at approximately 69500 and 27500. The latter is consistent with previous reports for the rat and mouse serum GHBPs based on nucleotide sequence analysis. The minor bands were essentially undetectable after storage at -25°C for several months, and an additional major band at M approximately 52500 appeared. The M(r) 69500 major protein contained N-linked carbohydrate, as determined by a reduction in molecular size by treatment with peptide N-glycosidase F. Binding of 125I-labelled GH was partially inhibited by co-incubation with 50 μg unlabelled pituitary-derived cGH/ml and excess unlabelled porcine GH as well as ovine prolactin, but not by bovine insulin. Non-specific binding of 125I-labelled GH by serum albumin was also observed. A comparison was made between these GHBPs and the hepatic GH receptor (e.g. molecular weight estimates, affinity for homologous versus heterologous GHs, cross-reactivity with prolactin, presence of N-linked carbohydrate). The origin and relationship among the various molecular weight species of GHBPs identified, and their potential role in regulation of the biological activity of GH in birds, remain to be determined.

AB - Multiple growth hormone (GH)-binding proteins (GHBPs) were identified in serum and plasma samples from domestic chickens and turkeys. Proteins were separated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis on 10% acrylamide, 2.7% bis discontinuous gels under reducing conditions and electrotransferred to nitrocellulose paper. Western blots were incubated with 125I-labelled recombinant chicken GH (cGH) or bovine GH and GHBPs visualized by means of autoradiography. In fresh samples (<2 h from collection to gel electrophoresis), multiple minor high M(r) bands were evident between approximately 72000 and 175000. Two major bands were observed at approximately 69500 and 27500. The latter is consistent with previous reports for the rat and mouse serum GHBPs based on nucleotide sequence analysis. The minor bands were essentially undetectable after storage at -25°C for several months, and an additional major band at M approximately 52500 appeared. The M(r) 69500 major protein contained N-linked carbohydrate, as determined by a reduction in molecular size by treatment with peptide N-glycosidase F. Binding of 125I-labelled GH was partially inhibited by co-incubation with 50 μg unlabelled pituitary-derived cGH/ml and excess unlabelled porcine GH as well as ovine prolactin, but not by bovine insulin. Non-specific binding of 125I-labelled GH by serum albumin was also observed. A comparison was made between these GHBPs and the hepatic GH receptor (e.g. molecular weight estimates, affinity for homologous versus heterologous GHs, cross-reactivity with prolactin, presence of N-linked carbohydrate). The origin and relationship among the various molecular weight species of GHBPs identified, and their potential role in regulation of the biological activity of GH in birds, remain to be determined.

UR - http://www.scopus.com/inward/record.url?scp=0025740204&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025740204&partnerID=8YFLogxK

M3 - Article

VL - 130

SP - 115

EP - 122

JO - Journal of Endocrinology

JF - Journal of Endocrinology

SN - 0022-0795

IS - 1

ER -