Document Detail


Binding Analysis of Ferritin with Heme Using α-Casein and Biotinylated-Hemin: Detection of Heme-Binding Capacity of Dpr Derived from Heme-Synthesis Deficient Streptococcus mutans.
MedLine Citation:
PMID:  23545463     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
Bacterial and mammalian ferritins are known to bind heme. The use of α-casein and biotinylated hemin could be applicable to detection of protein-bound heme and of proteins with heme-binding capacity, respectively. Although commercial horse spleen ferritin and purified horse spleen ferritin (L:H subunit ratio=4) bound to α-casein-coated plate, and this binding could be inhibited by hemin, recombinant iron-binding protein (rDpr), derived from heme-deficient Streptococcus mutans and expressed in Esherichia coli, did not bind to α-casein-coated plate. Both horse spleen ferritins bound to α-casein-immobilized beads. Commercial horse spleen ferritin and rDpr showed direct binding to hemin-agarose beads. After pre-incubation of commercial horse spleen ferritin or rDpr with biotinylated hemin, they showed indirect binding to avidin-immobilized beads through biotinylated hemin. These results demonstrate that α-casein is useful for detection of heme-binding ferritin, and both hemin-agarose and the combination of biotinylated hemin and avidin-beads are useful for the detection of heme-binding capacity of ferritin. In addition, this study also revealed that Dpr, decameric iron-binding protein, from heme-deficient cells binds heme.
Authors:
Ayako Mieno; Yuji Yamamoto; Yasunaga Yoshikawa; Kiyotaka Watanabe; Takao Mukai; Koichi Orino
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2013-4-02
Journal Detail:
Title:  The Journal of veterinary medical science / the Japanese Society of Veterinary Science     Volume:  -     ISSN:  1347-7439     ISO Abbreviation:  J. Vet. Med. Sci.     Publication Date:  2013 Apr 
Date Detail:
Created Date:  2013-4-2     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9105360     Medline TA:  J Vet Med Sci     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Affiliation:
Laboratory of Veterinary Biochemistry, School of Veterinary Medicine, Kitasato University.
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