Literature DB >> 568467

Solubilization of bovine corpus-luteum adenylate cyclase in lubrol-PX, triton X-100 or digitonin and the stabilizing effect of sodium fluoride present in the solubilization medium.

J L Young, D A Stansfield.   

Abstract

1. Adenylate cyclase activity of the washed 600g sediment of bovine corpus-luteum homogenate was solubilized by Lubrol-PX, Triton X-100 and digitonin. Digitonin was the least destructive of NaF-stimulated activity. 2. NaF, present in the solubilization medium together with MgSO4, increased the percentage yields of soluble activity from untreated 600g sediment and 600g sediment which had been preincubated with p[NH]ppG (guanosine 5'-[betagamma-imido]triphosphate). The stabilizing influence of NaF was most marked with digitonin. However, the highest specific activities of soluble enzyme were obtained with Lubrol-PX as solubilizing agent, since digitonin solubilized more membrane protein than does Lubrol-PX, and less of the activity of the digitonin-dispersed 600g sediment was recovered in the 105000g supernatant. 3. p[NH]ppG also has a stabilizing effect when present during the solubilization, but less so than NaF. 4. Both NaF and MgSO4 alone have a stabilizing effect during solubilization. The greatest amounts of soluble activity were obtained with both agents present in the solubilization medium, there being a synergistic effect.

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Year:  1978        PMID: 568467      PMCID: PMC1185859          DOI: 10.1042/bj1730919

Source DB:  PubMed          Journal:  Biochem J        ISSN: 0264-6021            Impact factor:   3.857


  11 in total

1.  Isolation of adenylate cyclase-free, beta-adrenergic receptor from turkey erythrocyte membranes by affinity chromatography.

Authors:  G Vauquelin; P Geynet; J Hanoune; A D Strosberg
Journal:  Proc Natl Acad Sci U S A       Date:  1977-09       Impact factor: 11.205

2.  Protein measurement with the Folin phenol reagent.

Authors:  O H LOWRY; N J ROSEBROUGH; A L FARR; R J RANDALL
Journal:  J Biol Chem       Date:  1951-11       Impact factor: 5.157

3.  Properties of adenylate cyclase solubilized from rat adrenal membranes: effects of ACTH and other stimulators on solubilization.

Authors:  D D Mahaffee; D A Ontjes
Journal:  J Cyclic Nucleotide Res       Date:  1977-10

4.  The vasopressin-sensitive adenylate cyclase of the rat renal medulla.

Authors:  E J Neer
Journal:  J Biol Chem       Date:  1973-07-10       Impact factor: 5.157

5.  Proceedings: Solubilization of adenylate cyclase of rat corpora lutes.

Authors:  G H Wilkinson; D A Stansfield
Journal:  Biochem Soc Trans       Date:  1974       Impact factor: 5.407

6.  Use of a competitive protein-binding assay for adenosine 3':5'-phosphate for the study of bovine corpus luteum adenylate cyclase.

Authors:  J L Young; D A Stansfield
Journal:  J Endocrinol       Date:  1977-04       Impact factor: 4.286

7.  Irreversible activation of adenylate cyclase of toad erythrocyte plasma membrane by 5'-guanylylimidodiphosphate.

Authors:  V Bennett; P Cuatrecasas
Journal:  J Membr Biol       Date:  1976       Impact factor: 1.843

8.  Properties of detergent-solubilized adenylate cyclase and gonadotropin receptors of testis and ovary.

Authors:  M L Dufau; A J Baukal; D Ryan; K J Catt
Journal:  Mol Cell Endocrinol       Date:  1977-02       Impact factor: 4.102

9.  Activation and inhibition of fat cell adenylate cyclase by fluoride.

Authors:  V C Manganiello; M Vaughan
Journal:  J Biol Chem       Date:  1976-10-25       Impact factor: 5.157

10.  Stabilization and solubilization of bovine corpus-luteum adenylate cyclase. The effects of guanosine triphosphate, guanosine 5'-[beta,gamma-imido]triphosphate, sodium fluoride and Tris/hydrochloric acid concentration on enzyme activity.

Authors:  J L Young; D A Stansfield
Journal:  Biochem J       Date:  1978-01-01       Impact factor: 3.857

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  2 in total

1.  Gel-filtration analysis of soluble adenylate cyclase from bovine corpus luteum.

Authors:  J L Young; D A Stansfield
Journal:  Biochem J       Date:  1978-11-01       Impact factor: 3.857

2.  Activation of adenylate cyclase in bovine corpus-luteum membranes by human choriogonadotropin, guanine nucleotides and NaF.

Authors:  N B Lydon; J L Young; D A Stansfield
Journal:  Biochem J       Date:  1981-09-15       Impact factor: 3.857

  2 in total

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