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Planta

, Volume 164, Issue 2, pp 272–277 | Cite as

Partial purification of an ethylene-binding site from Phaseolus vulgaris L. cotyledons

  • C. J. R. Thomas
  • A. R. Smith
  • M. A. Hall
Article

Abstract

The solubilised ethylene-binding site (EBS) of Phaseolus vulgaris L. cotyledons is an asymmetrical protein with a sedimentation coefficient of 2 S and a Stoke's radius of 6.1 nm (determined by ultracentrifugation on isokinetic gradients and gel-permeation chromatography, respectively). The molecular weight and frictional ratio were calculated as 52 000–60 000 and 2.37–2.48, respectively. The EBS has an isoelectric point at between pH 3–5, determined by isoelectric focussing and exhibits a negative charge at pH 8 during non-denaturing electrophoresis. The electrical charge on the EBS is shielded; the EBS does not bind to anion-exchange media under the experimental conditions reported here, is not precipitated by ammonium sulphate and does not precipitate at its isoelectric pH. The EBS preferentially partitions into detergent phases. The results indicate that the EBS is a hydrophobic protein complexed with detergent in aqueous solution. The techniques used to characterise the EBS also resulted in varying degress of purification.

Key words

Binding site (ethylene) Ethylene (binding site) Phaseolus (ethylene binding) 

Abbreviations

EBS

ethylene-binding site

Tricine

N-[2-hydroxy-1,1-bis(hydroxymethyl)-ethyl]glycine

Tris

2-amino-2-(hydroxymethyl)-1,3-propanediol

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Copyright information

© Springer-Verlag 1985

Authors and Affiliations

  • C. J. R. Thomas
    • 1
  • A. R. Smith
    • 2
  • M. A. Hall
    • 2
  1. 1.Biochemistry DepartmentNational Vegetable Research StationWellesbourne
  2. 2.Department of Botany and MicrobiologyUniversity College of WalesAberystwythUK

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