Document Detail


Proteolytic degradation of hemoglobin in the intestine of the human hookworm Necator americanus.
MedLine Citation:
PMID:  19434933     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Blood-feeding parasites use mechanistically distinct proteases to digest hemoglobin (Hb), often as multienzyme cooperative cascades. We investigated the roles played by 3 distinct proteases from adults of the human hookworm Necator americanus. The aspartic protease Na-APR-1 and the cysteine protease Na-CP-3 were expressed in catalytically active form in yeast, and the metalloprotease Na-MEP-1 was expressed in catalytically active form in baculovirus. Antibodies to all 3 proteases were used to immunolocalize each native enzyme to the intestine of adult N. americanus. Recombinant Na-APR-1 cleaved intact human Hb. In contrast, Na-CP-3 and Na-MEP-1 could not cleave Hb but instead cleaved globin fragments that had been hydrolyzed by Na-APR-1, implying an ordered process of hemoglobinolysis. Seventy-four cleavage sites within Hb alpha- and beta-chains were characterized after digestion with all 3 proteases. All of the proteases demonstrated promiscuous subsite specificities within Hb; noteworthy preferences included aromatic and hydrophobic P1 residues and hydrophobic P1' residues for Na-APR-1 and hydrophobic P1 residues for Na-MEP-1. We conclude that Hb digestion in N. americanus involves a network of distinct proteases, some of which act in an ordered fashion, providing a potential mechanism by which some of these hemoglobinases exert their efficacy as recombinant vaccines against hookworm infection.
Authors:
Najju Ranjit; Bin Zhan; Brett Hamilton; Deborah Stenzel; Jonathan Lowther; Mark Pearson; Jeffrey Gorman; Peter Hotez; Alex Loukas
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The Journal of infectious diseases     Volume:  199     ISSN:  0022-1899     ISO Abbreviation:  J. Infect. Dis.     Publication Date:  2009 Mar 
Date Detail:
Created Date:  2009-05-12     Completed Date:  2009-05-29     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  0413675     Medline TA:  J Infect Dis     Country:  United States    
Other Details:
Languages:  eng     Pagination:  904-12     Citation Subset:  AIM; IM    
Affiliation:
Helminth Biology Laboratory, Division of Infectious Diseases, Queensland Institute of Medical Research, Australia.
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Animals
Aspartic Acid Endopeptidases / metabolism
Catalysis
Cloning, Molecular
Hemoglobins / chemistry,  genetics,  metabolism*
Humans
Hydrolysis
Kinetics
Molecular Sequence Data
Necator americanus / enzymology,  metabolism*
Necatoriasis / immunology,  metabolism*
Peptide Fragments / chemistry,  metabolism
Peptide Hydrolases / metabolism
Plasmodium falciparum / metabolism
Recombinant Proteins / metabolism
Chemical
Reg. No./Substance:
0/Hemoglobins; 0/Peptide Fragments; 0/Recombinant Proteins; 137597-31-0/alpha(A) globin; EC 3.4.-/Peptide Hydrolases; EC 3.4.23.-/Aspartic Acid Endopeptidases

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


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