Resolution and activity of adenylate cyclase components in a zwitterionic cholate derivative [3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate]

Alan J. Bitonti, Joel Moss, Leonard Hjelmeland, Leonard M Hjelmeland

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

Bovine brain adenylate cyclase was solubilized with 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS), sodium cholate, sodium deoxycholate, or these detergents plus (NH4)2SO4. The specific activity of the extract obtained with 13 mM CHAPS alone was several times those of the other detergent extracts with or without (NH4)2SO4. After solubilization with 13 mM CHAPS, gel filtration completely separated the catalytic unit (C) from the guanine nucleotide binding protein (G/F). C activity when assayed with 5 mM Mn2+ was 5 times that assayed with 10 mM Mg2+ and was unresponsive to GPP(NH)P. C activity was increased ∼150% by GPP(NH)P in the presence of G/F extracted from human erythrocyte ghosts and ∼100% by Ca2+ plus calmodulin in assays with Mg2+. On gel filtration and/or density gradient centrifugation, the physical properties of C from brain or AC- cells and G/F from bovine or pig erythrocytes in CHAPs were similar to those observed in other detergents. It appears that the use of CHAPS for solubilization of adenylate cyclase and separation of C and G/F may well prove advantageous in studies of the molecular interactions between the protein subunits and activators of the enzyme as well as for the initial purification of C.

Original languageEnglish (US)
Pages (from-to)3650-3653
Number of pages4
JournalBiochemistry
Volume21
Issue number15
StatePublished - 1982
Externally publishedYes

Fingerprint

Cholates
Adenylyl Cyclases
Detergents
Derivatives
Gel Chromatography
Brain
Gels
Enzyme Activators
Sodium Cholate
Gastrin-Secreting Cells
Deoxycholic Acid
Guanine Nucleotides
Density Gradient Centrifugation
Molecular interactions
Centrifugation
Protein Subunits
Erythrocyte Membrane
Calmodulin
Purification
Assays

ASJC Scopus subject areas

  • Biochemistry

Cite this

Resolution and activity of adenylate cyclase components in a zwitterionic cholate derivative [3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate]. / Bitonti, Alan J.; Moss, Joel; Hjelmeland, Leonard; Hjelmeland, Leonard M.

In: Biochemistry, Vol. 21, No. 15, 1982, p. 3650-3653.

Research output: Contribution to journalArticle

@article{82b49b6e588d444bb529bdea2c76e6f0,
title = "Resolution and activity of adenylate cyclase components in a zwitterionic cholate derivative [3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate]",
abstract = "Bovine brain adenylate cyclase was solubilized with 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS), sodium cholate, sodium deoxycholate, or these detergents plus (NH4)2SO4. The specific activity of the extract obtained with 13 mM CHAPS alone was several times those of the other detergent extracts with or without (NH4)2SO4. After solubilization with 13 mM CHAPS, gel filtration completely separated the catalytic unit (C) from the guanine nucleotide binding protein (G/F). C activity when assayed with 5 mM Mn2+ was 5 times that assayed with 10 mM Mg2+ and was unresponsive to GPP(NH)P. C activity was increased ∼150{\%} by GPP(NH)P in the presence of G/F extracted from human erythrocyte ghosts and ∼100{\%} by Ca2+ plus calmodulin in assays with Mg2+. On gel filtration and/or density gradient centrifugation, the physical properties of C from brain or AC- cells and G/F from bovine or pig erythrocytes in CHAPs were similar to those observed in other detergents. It appears that the use of CHAPS for solubilization of adenylate cyclase and separation of C and G/F may well prove advantageous in studies of the molecular interactions between the protein subunits and activators of the enzyme as well as for the initial purification of C.",
author = "Bitonti, {Alan J.} and Joel Moss and Leonard Hjelmeland and Hjelmeland, {Leonard M}",
year = "1982",
language = "English (US)",
volume = "21",
pages = "3650--3653",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "15",

}

TY - JOUR

T1 - Resolution and activity of adenylate cyclase components in a zwitterionic cholate derivative [3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate]

AU - Bitonti, Alan J.

AU - Moss, Joel

AU - Hjelmeland, Leonard

AU - Hjelmeland, Leonard M

PY - 1982

Y1 - 1982

N2 - Bovine brain adenylate cyclase was solubilized with 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS), sodium cholate, sodium deoxycholate, or these detergents plus (NH4)2SO4. The specific activity of the extract obtained with 13 mM CHAPS alone was several times those of the other detergent extracts with or without (NH4)2SO4. After solubilization with 13 mM CHAPS, gel filtration completely separated the catalytic unit (C) from the guanine nucleotide binding protein (G/F). C activity when assayed with 5 mM Mn2+ was 5 times that assayed with 10 mM Mg2+ and was unresponsive to GPP(NH)P. C activity was increased ∼150% by GPP(NH)P in the presence of G/F extracted from human erythrocyte ghosts and ∼100% by Ca2+ plus calmodulin in assays with Mg2+. On gel filtration and/or density gradient centrifugation, the physical properties of C from brain or AC- cells and G/F from bovine or pig erythrocytes in CHAPs were similar to those observed in other detergents. It appears that the use of CHAPS for solubilization of adenylate cyclase and separation of C and G/F may well prove advantageous in studies of the molecular interactions between the protein subunits and activators of the enzyme as well as for the initial purification of C.

AB - Bovine brain adenylate cyclase was solubilized with 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS), sodium cholate, sodium deoxycholate, or these detergents plus (NH4)2SO4. The specific activity of the extract obtained with 13 mM CHAPS alone was several times those of the other detergent extracts with or without (NH4)2SO4. After solubilization with 13 mM CHAPS, gel filtration completely separated the catalytic unit (C) from the guanine nucleotide binding protein (G/F). C activity when assayed with 5 mM Mn2+ was 5 times that assayed with 10 mM Mg2+ and was unresponsive to GPP(NH)P. C activity was increased ∼150% by GPP(NH)P in the presence of G/F extracted from human erythrocyte ghosts and ∼100% by Ca2+ plus calmodulin in assays with Mg2+. On gel filtration and/or density gradient centrifugation, the physical properties of C from brain or AC- cells and G/F from bovine or pig erythrocytes in CHAPs were similar to those observed in other detergents. It appears that the use of CHAPS for solubilization of adenylate cyclase and separation of C and G/F may well prove advantageous in studies of the molecular interactions between the protein subunits and activators of the enzyme as well as for the initial purification of C.

UR - http://www.scopus.com/inward/record.url?scp=0020487507&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0020487507&partnerID=8YFLogxK

M3 - Article

VL - 21

SP - 3650

EP - 3653

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 15

ER -