Document Detail

Characterization of cucurbita maxima phloem serpin-1 (CmPS-1). A developmentally regulated elastase inhibitor.
MedLine Citation:
PMID:  10960478     Owner:  NLM     Status:  MEDLINE    
We report on the molecular, biochemical, and functional characterization of Cucurbita maxima phloem serpin-1 (CmPS-1), a novel 42-kDa serine proteinase inhibitor that is developmentally regulated and has anti-elastase properties. CmPS-1 was purified to near homogeneity from C. maxima (pumpkin) phloem exudate and, based on microsequence analysis, the cDNA encoding CmPS-1 was cloned. The association rate constant (k(a)) of phloem-purified and recombinant His(6)-tagged CmPS-1 for elastase was 3.5 +/- 1.6 x 10(5) and 2.7 +/- 0.4 x 10(5) m(-)(1) s(-)(1), respectively. The fraction of complex-forming CmPS-1, X(inh), was estimated at 79%. CmPS-1 displayed no detectable inhibitory properties against chymotrypsin, trypsin, or thrombin. The elastase cleavage sites within the reactive center loop of CmPS-1 were determined to be Val(347)-Gly(348) and Val(350)-Ser(351) with a 3:2 molar ratio. In vivo feeding assays conducted with the piercing-sucking aphid, Myzus persicae, established a close correlation between the developmentally regulated increase in CmPS-1 within the phloem sap and the reduced ability of these insects to survive and reproduce on C. maxima. However, in vitro feeding experiments, using purified phloem CmPS-1, failed to demonstrate a direct effect on aphid survival. Likely roles of this novel phloem serpin in defense against insects/pathogens are discussed.
B C Yoo; K Aoki; Y Xiang; L R Campbell; R J Hull; B Xoconostle-Cázares; J Monzer; J Y Lee; D E Ullman; W J Lucas
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  275     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  2000 Nov 
Date Detail:
Created Date:  2000-11-27     Completed Date:  2001-01-04     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  35122-8     Citation Subset:  IM    
Department of Entomolgy, College of Agriculture and Environmental Sciences, University of California, Davis, California 95616, USA.
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MeSH Terms
Amino Acid Sequence
Binding Sites
Blotting, Western
Chymotrypsin / pharmacology
Cloning, Molecular
Cucurbitaceae / chemistry
DNA, Complementary / metabolism
Electrophoresis, Polyacrylamide Gel
Glycine / chemistry
Molecular Sequence Data
Polymerase Chain Reaction
Protein Binding
Recombinant Proteins / chemistry,  metabolism
Sequence Analysis, DNA
Sequence Homology, Amino Acid
Serine / chemistry
Serine Proteinase Inhibitors / chemistry*
Serpins / chemistry*,  genetics,  metabolism
Thrombin / pharmacology
Time Factors
Trypsin / pharmacology
Valine / chemistry
Reg. No./Substance:
0/DNA, Complementary; 0/Recombinant Proteins; 0/Serine Proteinase Inhibitors; 0/Serpins; 0/phloem serpin 1; 56-40-6/Glycine; 56-45-1/Serine; 7004-03-7/Valine; EC; EC; EC

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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