Document Detail


Characterization of cucurbita maxima phloem serpin-1 (CmPS-1). A developmentally regulated elastase inhibitor.
MedLine Citation:
PMID:  10960478     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
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
Related Documents :
11468408 - Coexpression, copurification, crystallization and preliminary x-ray analysis of a compl...
21578318 - 2-chloro-7-methyl-quinoline-3-carbaldehyde.
19874048 - Identification of the rate-determining step of trna-guanine transglycosylase from esche...
10987918 - Probing the stereochemistry of e. coli 3-deoxy-d-arabino-heptulosonate 7-phosphate synt...
16584778 - Production and characterization of a polyclonal antibody for os(ii) and ru(ii) polypyri...
23747378 - Synthesis of a new n-substituted bis-benzimidazolyl diamide ligand and its trinuclear c...
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    
Affiliation:
Department of Entomolgy, College of Agriculture and Environmental Sciences, University of California, Davis, California 95616, USA.
Data Bank Information
Bank Name/Acc. No.:
GENBANK/AF284038
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Animals
Aphids
Binding Sites
Blotting, Western
Chymotrypsin / pharmacology
Cloning, Molecular
Cucurbitaceae / chemistry
DNA, Complementary / metabolism
Electrophoresis, Polyacrylamide Gel
Glycine / chemistry
Kinetics
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
Chemical
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 3.4.21.1/Chymotrypsin; EC 3.4.21.4/Trypsin; EC 3.4.21.5/Thrombin

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


Previous Document:  CTP:2,3-di-O-geranylgeranyl-sn-glycero-1-phosphate cytidyltransferase in the methanogenic archaeon M...
Next Document:  Mitogen-activated protein kinase plays an essential role in the erythropoietin-dependent proliferati...