PMID- 9792913
OWN - NLM
STAT- MEDLINE
DCOM- 19990211
LR  - 20190512
IS  - 0021-924X (Print)
IS  - 0021-924X (Linking)
VI  - 124
IP  - 5
DP  - 1998 Nov
TI  - Molecular cloning, functional expression, and mutagenesis of cDNA encoding a 
      cysteine proteinase inhibitor from sunflower seeds.
PG  - 911-6
AB  - Sunflower cystatin Scb differs from other phytocystatins in that it is a highly 
      basic protein with a pI value of 9.6 and includes six additional amino acids 
      (Arg30-Leu-Gln-Arg-Thr34, Thr37) in the middle region as compared with other 
      phytocystatins [Kouzuma et al. (1996) J. Biochem. 119, 1106-1113]. We identified 
      and sequenced a complete cDNA encoding the Scb; the cDNA of Scb consists of 645 
      nucleotides and includes an open reading frame encoding a polypeptide of 123 
      amino acids. On the basis of these findings, Scb appears to be synthesized as a 
      prepeptide consisting of a signal sequence of 22 amino acids and a mature protein 
      of 101 amino acids. A recombinant Scb (rScb) was produced by expression in 
      Escherichia coli and purified by gel filtration on Sephacryl S-200 followed by 
      ion-exchange column chromatography on a S-Sepharose column. rScb exhibited almost 
      the same inhibitory activity toward papain as the authentic Scb did, but its 
      inhibition profile toward cathepsins B, L, and H was slightly different. Scb 
      mutant proteins, in which selected N-terminal residues or the additional amino 
      acids were deleted, were subsequently constructed and characterized with respect 
      to their inhibitory activities toward papain. The result revealed that the 
      additional sequence (Arg30-Leu-Gln-Arg-Thr34) in Scb is not essential for 
      papain-inhibitory activity, while the N-terminal amino acids (Ile1-Pro2) as well 
      as the N-terminal glycine residues Gly3 and/or Gly4 play an important role in 
      manifesting the inhibitory activity toward papain.
FAU - Doi-Kawano, K
AU  - Doi-Kawano K
AD  - Laboratory of Biochemistry, Faculty of Agriculture, Kyushu University, 
      Higashi-ku, Fukuoka, 812-8581, Japan.
FAU - Kouzuma, Y
AU  - Kouzuma Y
FAU - Yamasaki, N
AU  - Yamasaki N
FAU - Kimura, M
AU  - Kimura M
LA  - eng
PT  - Journal Article
PL  - England
TA  - J Biochem
JT  - Journal of biochemistry
JID - 0376600
RN  - 0 (Cystatins)
RN  - 0 (Cysteine Proteinase Inhibitors)
RN  - 0 (DNA, Complementary)
RN  - 0 (Plant Proteins)
RN  - 0 (Scb protein, Helianthus annuus)
SB  - IM
MH  - Amino Acid Sequence
MH  - Base Sequence
MH  - Cloning, Molecular
MH  - Cystatins/*genetics
MH  - Cysteine Proteinase Inhibitors/*genetics
MH  - DNA, Complementary
MH  - Escherichia coli/genetics
MH  - Helianthus/*chemistry
MH  - Molecular Sequence Data
MH  - Mutagenesis, Site-Directed
MH  - *Plant Proteins
MH  - Seeds/chemistry
EDAT- 1998/10/30 00:00
MHDA- 1998/10/30 00:01
CRDT- 1998/10/30 00:00
PHST- 1998/10/30 00:00 [pubmed]
PHST- 1998/10/30 00:01 [medline]
PHST- 1998/10/30 00:00 [entrez]
AID - 10.1093/oxfordjournals.jbchem.a022207 [doi]
PST - ppublish
SO  - J Biochem. 1998 Nov;124(5):911-6. doi: 10.1093/oxfordjournals.jbchem.a022207.