Isolation and characterization of biologically active and inactive cholecystokinin-octapeptides from human brain

J. R. Reeve, V. E. Eysselein, J. H. Walsh, H. Sankaran, Clifford Deveney, W. W. Tourtellotte, C. Miller, J. E. Shively

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Cholecystokinin-like immunoreactivity (CCK-LI) in 0.9 kg human brain was extracted by 2% trifluoroacetic acid at 4°C. Sephadex G50 gel filtration of crude extract revealed one main molecular form of CCK, detected by a carboxy-terminal antibody (5135), that eluted in the position of CCK8. When the CCK-LI in the extract was purified by affinity chromatography using another carboxyl-terminal CCK antibody followed by several steps of reverse phase high pressure liquid chromatography (HPLC), a component was isolated that was found by sequence analysis to be identical to the carboxyl-terminal CCK-octapeptide of porcine CCK33, isolated from intestinal mucosa, and to CCK-octapeptide, isolated from sheep brain. This component possessed comparable biological potencies to synthetic sulfated CCK8 in eliciting amylase release and in competitively displacing radioiodinated CCK33 from isolated mouse pancreatic acini. Furthermore, it exhibited a similar binding characteristic to CCK8 in binding to specific receptors on mouse brain cortical particulate preparations. On high pressure liquid chromatography another minor, earlier eluting immunoreactive peak was observed, which had the same amino acid composition and sequence as CCK8. These findings suggested that this material was oxidized CCK8. This earlier eluting component, exhibiting CCK8-like immunoreactivity, did not induce amylase release from acini and had no or minimal effect in inhibiting tracer CCK33 binding to receptors on isolated acini or on mouse brain cortical particulate preparations at the concentrations tested.

Original languageEnglish (US)
Pages (from-to)959-966
Number of pages8
JournalPeptides
Volume5
Issue number5
DOIs
StatePublished - 1984
Externally publishedYes

Fingerprint

Sincalide
Brain
High pressure liquid chromatography
Cholecystokinin
Amylases
High Pressure Liquid Chromatography
Affinity chromatography
Trifluoroacetic Acid
Antibodies
Reverse-Phase Chromatography
Intestinal Mucosa
Complex Mixtures
Affinity Chromatography
Gel Chromatography
Sequence Analysis
Amino Acid Sequence
Sheep
Swine
Amino Acids
Chemical analysis

Keywords

  • Amino acid sequence
  • CCK biological activity
  • Cholecystokinin
  • Human brain peptides

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology
  • Physiology
  • Cellular and Molecular Neuroscience

Cite this

Reeve, J. R., Eysselein, V. E., Walsh, J. H., Sankaran, H., Deveney, C., Tourtellotte, W. W., ... Shively, J. E. (1984). Isolation and characterization of biologically active and inactive cholecystokinin-octapeptides from human brain. Peptides, 5(5), 959-966. https://doi.org/10.1016/0196-9781(84)90123-2

Isolation and characterization of biologically active and inactive cholecystokinin-octapeptides from human brain. / Reeve, J. R.; Eysselein, V. E.; Walsh, J. H.; Sankaran, H.; Deveney, Clifford; Tourtellotte, W. W.; Miller, C.; Shively, J. E.

In: Peptides, Vol. 5, No. 5, 1984, p. 959-966.

Research output: Contribution to journalArticle

Reeve, JR, Eysselein, VE, Walsh, JH, Sankaran, H, Deveney, C, Tourtellotte, WW, Miller, C & Shively, JE 1984, 'Isolation and characterization of biologically active and inactive cholecystokinin-octapeptides from human brain', Peptides, vol. 5, no. 5, pp. 959-966. https://doi.org/10.1016/0196-9781(84)90123-2
Reeve, J. R. ; Eysselein, V. E. ; Walsh, J. H. ; Sankaran, H. ; Deveney, Clifford ; Tourtellotte, W. W. ; Miller, C. ; Shively, J. E. / Isolation and characterization of biologically active and inactive cholecystokinin-octapeptides from human brain. In: Peptides. 1984 ; Vol. 5, No. 5. pp. 959-966.
@article{94e580a1f92c40ee8c0ad7061af0ef46,
title = "Isolation and characterization of biologically active and inactive cholecystokinin-octapeptides from human brain",
abstract = "Cholecystokinin-like immunoreactivity (CCK-LI) in 0.9 kg human brain was extracted by 2{\%} trifluoroacetic acid at 4°C. Sephadex G50 gel filtration of crude extract revealed one main molecular form of CCK, detected by a carboxy-terminal antibody (5135), that eluted in the position of CCK8. When the CCK-LI in the extract was purified by affinity chromatography using another carboxyl-terminal CCK antibody followed by several steps of reverse phase high pressure liquid chromatography (HPLC), a component was isolated that was found by sequence analysis to be identical to the carboxyl-terminal CCK-octapeptide of porcine CCK33, isolated from intestinal mucosa, and to CCK-octapeptide, isolated from sheep brain. This component possessed comparable biological potencies to synthetic sulfated CCK8 in eliciting amylase release and in competitively displacing radioiodinated CCK33 from isolated mouse pancreatic acini. Furthermore, it exhibited a similar binding characteristic to CCK8 in binding to specific receptors on mouse brain cortical particulate preparations. On high pressure liquid chromatography another minor, earlier eluting immunoreactive peak was observed, which had the same amino acid composition and sequence as CCK8. These findings suggested that this material was oxidized CCK8. This earlier eluting component, exhibiting CCK8-like immunoreactivity, did not induce amylase release from acini and had no or minimal effect in inhibiting tracer CCK33 binding to receptors on isolated acini or on mouse brain cortical particulate preparations at the concentrations tested.",
keywords = "Amino acid sequence, CCK biological activity, Cholecystokinin, Human brain peptides",
author = "Reeve, {J. R.} and Eysselein, {V. E.} and Walsh, {J. H.} and H. Sankaran and Clifford Deveney and Tourtellotte, {W. W.} and C. Miller and Shively, {J. E.}",
year = "1984",
doi = "10.1016/0196-9781(84)90123-2",
language = "English (US)",
volume = "5",
pages = "959--966",
journal = "Peptides",
issn = "0196-9781",
publisher = "Elsevier Inc.",
number = "5",

}

TY - JOUR

T1 - Isolation and characterization of biologically active and inactive cholecystokinin-octapeptides from human brain

AU - Reeve, J. R.

AU - Eysselein, V. E.

AU - Walsh, J. H.

AU - Sankaran, H.

AU - Deveney, Clifford

AU - Tourtellotte, W. W.

AU - Miller, C.

AU - Shively, J. E.

PY - 1984

Y1 - 1984

N2 - Cholecystokinin-like immunoreactivity (CCK-LI) in 0.9 kg human brain was extracted by 2% trifluoroacetic acid at 4°C. Sephadex G50 gel filtration of crude extract revealed one main molecular form of CCK, detected by a carboxy-terminal antibody (5135), that eluted in the position of CCK8. When the CCK-LI in the extract was purified by affinity chromatography using another carboxyl-terminal CCK antibody followed by several steps of reverse phase high pressure liquid chromatography (HPLC), a component was isolated that was found by sequence analysis to be identical to the carboxyl-terminal CCK-octapeptide of porcine CCK33, isolated from intestinal mucosa, and to CCK-octapeptide, isolated from sheep brain. This component possessed comparable biological potencies to synthetic sulfated CCK8 in eliciting amylase release and in competitively displacing radioiodinated CCK33 from isolated mouse pancreatic acini. Furthermore, it exhibited a similar binding characteristic to CCK8 in binding to specific receptors on mouse brain cortical particulate preparations. On high pressure liquid chromatography another minor, earlier eluting immunoreactive peak was observed, which had the same amino acid composition and sequence as CCK8. These findings suggested that this material was oxidized CCK8. This earlier eluting component, exhibiting CCK8-like immunoreactivity, did not induce amylase release from acini and had no or minimal effect in inhibiting tracer CCK33 binding to receptors on isolated acini or on mouse brain cortical particulate preparations at the concentrations tested.

AB - Cholecystokinin-like immunoreactivity (CCK-LI) in 0.9 kg human brain was extracted by 2% trifluoroacetic acid at 4°C. Sephadex G50 gel filtration of crude extract revealed one main molecular form of CCK, detected by a carboxy-terminal antibody (5135), that eluted in the position of CCK8. When the CCK-LI in the extract was purified by affinity chromatography using another carboxyl-terminal CCK antibody followed by several steps of reverse phase high pressure liquid chromatography (HPLC), a component was isolated that was found by sequence analysis to be identical to the carboxyl-terminal CCK-octapeptide of porcine CCK33, isolated from intestinal mucosa, and to CCK-octapeptide, isolated from sheep brain. This component possessed comparable biological potencies to synthetic sulfated CCK8 in eliciting amylase release and in competitively displacing radioiodinated CCK33 from isolated mouse pancreatic acini. Furthermore, it exhibited a similar binding characteristic to CCK8 in binding to specific receptors on mouse brain cortical particulate preparations. On high pressure liquid chromatography another minor, earlier eluting immunoreactive peak was observed, which had the same amino acid composition and sequence as CCK8. These findings suggested that this material was oxidized CCK8. This earlier eluting component, exhibiting CCK8-like immunoreactivity, did not induce amylase release from acini and had no or minimal effect in inhibiting tracer CCK33 binding to receptors on isolated acini or on mouse brain cortical particulate preparations at the concentrations tested.

KW - Amino acid sequence

KW - CCK biological activity

KW - Cholecystokinin

KW - Human brain peptides

UR - http://www.scopus.com/inward/record.url?scp=0021723371&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0021723371&partnerID=8YFLogxK

U2 - 10.1016/0196-9781(84)90123-2

DO - 10.1016/0196-9781(84)90123-2

M3 - Article

VL - 5

SP - 959

EP - 966

JO - Peptides

JF - Peptides

SN - 0196-9781

IS - 5

ER -