Expression and function of a cysteine proteinase cDNA from sweet potato (Ipomoea batatas [L.] Lam 'Tainong 57') storage roots

Dong Jiann Huang, Hsien Jung Chen, Wen C. Hou, Tzeng E. Chen, Wei Y. Hsu, Yaw Huei Lin

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Cysteine proteinase (CPR) cDNA clone (SPCPRPP) of sweet potato (Ipomoea batatas [L.] Lam 'Tainong 57') storage roots were isolated by differential display. The open reading frame in this cDNA encodes a pre-proprotein of 371 amino acids with conserved catalytic amino acids of papain. Examination of the expression patterns in sweet potato by Northern blot analyses revealed that the transcripts of SPCPRPP were specifically induced in the storage roots. Recombinant SPCPRPP protein overproduced in Escherichia coli (M15) was purified by Ni2+-chelated affinity chromatography. Active recombinant SPCPRPP protein was able to digest the 22 kDa sweet potato trypsin inhibitor (TI) protein when the latter was reduced by DTT (dithiothreitol) or NTS (NADPH/thioredoxin system). A smaller peptide (14 kDa) was obtained as a digestion product. These results suggest that CPR is responsible for initiation of degradation and re-mobilization of stored 22 kDa TI during sprouting of SP storage roots after the reduction of 22 kDa TI by the NTS.

Original languageEnglish
Pages (from-to)423-431
Number of pages9
JournalPlant Science
Volume169
Issue number2
DOIs
Publication statusPublished - Aug 2005

Fingerprint

Ipomoea batatas
Trypsin Inhibitors
Cysteine Proteases
cysteine proteinases
trypsin inhibitors
sweet potatoes
Thioredoxins
Complementary DNA
NADP
Recombinant Proteins
recombinant proteins
NADP (coenzyme)
Affinity chromatography
Amino Acids
amino acids
Papain
dithiothreitol
papain
Dithiothreitol
affinity chromatography

Keywords

  • cDNA sequence
  • Cysteine proteinase
  • Gene expression
  • Recombinant protein
  • Sweet potato
  • Trypsin inhibitors

ASJC Scopus subject areas

  • Plant Science
  • Biochemistry
  • Biotechnology

Cite this

Expression and function of a cysteine proteinase cDNA from sweet potato (Ipomoea batatas [L.] Lam 'Tainong 57') storage roots. / Huang, Dong Jiann; Chen, Hsien Jung; Hou, Wen C.; Chen, Tzeng E.; Hsu, Wei Y.; Lin, Yaw Huei.

In: Plant Science, Vol. 169, No. 2, 08.2005, p. 423-431.

Research output: Contribution to journalArticle

Huang, Dong Jiann ; Chen, Hsien Jung ; Hou, Wen C. ; Chen, Tzeng E. ; Hsu, Wei Y. ; Lin, Yaw Huei. / Expression and function of a cysteine proteinase cDNA from sweet potato (Ipomoea batatas [L.] Lam 'Tainong 57') storage roots. In: Plant Science. 2005 ; Vol. 169, No. 2. pp. 423-431.
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