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Biologia Plantarum

, 26:99 | Cite as

Detection and evaluation of serine proteinase by affinity chromatography on immobilized-aprotinin inRicinus communia

  • Tsuneo Watanabe
  • Noriaki Kondo
  • Kazutaka Kano
Original Papers

Abstract

Neutral proteinase was found in the leaves ofRicinus communie as assayed with α-casein and H-D-Val-Leu-Lys-pNA as substrates. The enzyme is maximally active at pH around 7.4. A selective adsorbent for serine proteinase was prepared by attaching aprotinin to aminoalkyl-porous glass.

When partially purified leaf proteinase was passed through a column containing this adsorbent, the proteinase activity present was bound to the porous glass. The proteinase eluted at IM NaCl was inhibited by aprotinin, leupeptin, DFP, phenylmethylsulfonyl fluoride (PMSF) and serine proteinase inhibitor fromR, communis leaves, whereas pepstatin, EDTA, EGTA, and DTT had no effect on the enzyme. This inhibition profile suggests the leaf proteinase is a neutral proteinase, such as a serine proteinase.

Keywords

Serine Proteinase Proteinase Activity Affinity Chromatography Plasmin Porous Glass 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Abbreviations used

DTT

dithiothreitol

DFP

diisopropylfluorophosphate

PMSF

phenylmethylsulfonylfluoride

PEG-6000

polyethylene glycol-6000

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Copyright information

© Academia 1984

Authors and Affiliations

  • Tsuneo Watanabe
    • 1
  • Noriaki Kondo
    • 1
  • Kazutaka Kano
    • 2
  1. 1.Laboratory of Biochemistry and Physiology, Division of Environmental BiologyNational Institute for Environmental StudiesIbarakiJapan
  2. 2.Department of Physiological Chemistry and Nutrition Faculty of MedicineUniversity of TokyoTokyoJapan

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