A probe for capture and Fe3+-induced conformational change of lactoferrin selected from phage displayed peptide libraries
(2004) In Journal of Dairy Science 87(10). p.3247-3255- Abstract
- Linear pentadecamer and cyclic hexamer peptide phage libraries were used to isolate phage clones with binding affinity toward lactoferrins purified from human and bovine milk. Phage clones with high specificity toward lactoferrin were selected with different binding strengths depending on the sequence of the peptide displayed by the phage. Phages coated to a microtiterplate were able to capture lactoferrin from crude milk samples without prior treatment. One of the selected sequences, EGKQRR, failed to bind to lactoferrin. In contrast, a branched tree-peptide bearing 4 EGKQRR sequences did bind to lactoferrin (Kdsimilar to29 muM) and was also capable of inhibiting the binding of the phage to lactoferrin (IC(50)similar to17 muM), indicating... (More)
- Linear pentadecamer and cyclic hexamer peptide phage libraries were used to isolate phage clones with binding affinity toward lactoferrins purified from human and bovine milk. Phage clones with high specificity toward lactoferrin were selected with different binding strengths depending on the sequence of the peptide displayed by the phage. Phages coated to a microtiterplate were able to capture lactoferrin from crude milk samples without prior treatment. One of the selected sequences, EGKQRR, failed to bind to lactoferrin. In contrast, a branched tree-peptide bearing 4 EGKQRR sequences did bind to lactoferrin (Kdsimilar to29 muM) and was also capable of inhibiting the binding of the phage to lactoferrin (IC(50)similar to17 muM), indicating that avidity was important. Unexpectedly, the affinity of the phage for lactoferrin was influenced by the amount of bound Fe3+, with a much lower affinity when lactoferrin was saturated with Fe3+ as compared with the iron-depleted or partially saturated (natural) lactoferrin. As the phage does not bind to the Fe3+-binding site, the difference in binding affinity is due to differences in conformation of lactoferrin induced by Fe3+. These results demonstrate that avidity or multipoint attachment and Fe3+-induced conformational changes play an important role in the binding of the selected phage to lactoferrin. Thus, we could demonstrate that, by the use of selected phage clones, we are able not only to detect lactoferrin, but also to capture lactoferrin from crude milk samples. Furthermore, the extent of phage binding provides additional information about the iron content and the concomitant conformation of lactoferrin. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/140580
- author
- Noppe, W ; Vanhoorelbeke, K ; Galaev, Igor LU ; Mattiasson, Bo LU and Deckmyn, H
- organization
- publishing date
- 2004
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Journal of Dairy Science
- volume
- 87
- issue
- 10
- pages
- 3247 - 3255
- publisher
- American Dairy Science Association
- external identifiers
-
- wos:000224011800016
- pmid:15377604
- scopus:6944254411
- ISSN
- 1525-3198
- language
- English
- LU publication?
- yes
- id
- c4b5a003-c536-4c92-958c-4b85450f900d (old id 140580)
- alternative location
- http://jds.fass.org/cgi/content/abstract/87/10/3247
- http://www.dairy-science.org/cgi/reprint/87/10/3247
- date added to LUP
- 2016-04-01 11:46:22
- date last changed
- 2022-03-28 02:53:37
@article{c4b5a003-c536-4c92-958c-4b85450f900d,
abstract = {{Linear pentadecamer and cyclic hexamer peptide phage libraries were used to isolate phage clones with binding affinity toward lactoferrins purified from human and bovine milk. Phage clones with high specificity toward lactoferrin were selected with different binding strengths depending on the sequence of the peptide displayed by the phage. Phages coated to a microtiterplate were able to capture lactoferrin from crude milk samples without prior treatment. One of the selected sequences, EGKQRR, failed to bind to lactoferrin. In contrast, a branched tree-peptide bearing 4 EGKQRR sequences did bind to lactoferrin (Kdsimilar to29 muM) and was also capable of inhibiting the binding of the phage to lactoferrin (IC(50)similar to17 muM), indicating that avidity was important. Unexpectedly, the affinity of the phage for lactoferrin was influenced by the amount of bound Fe3+, with a much lower affinity when lactoferrin was saturated with Fe3+ as compared with the iron-depleted or partially saturated (natural) lactoferrin. As the phage does not bind to the Fe3+-binding site, the difference in binding affinity is due to differences in conformation of lactoferrin induced by Fe3+. These results demonstrate that avidity or multipoint attachment and Fe3+-induced conformational changes play an important role in the binding of the selected phage to lactoferrin. Thus, we could demonstrate that, by the use of selected phage clones, we are able not only to detect lactoferrin, but also to capture lactoferrin from crude milk samples. Furthermore, the extent of phage binding provides additional information about the iron content and the concomitant conformation of lactoferrin.}},
author = {{Noppe, W and Vanhoorelbeke, K and Galaev, Igor and Mattiasson, Bo and Deckmyn, H}},
issn = {{1525-3198}},
language = {{eng}},
number = {{10}},
pages = {{3247--3255}},
publisher = {{American Dairy Science Association}},
series = {{Journal of Dairy Science}},
title = {{A probe for capture and Fe3+-induced conformational change of lactoferrin selected from phage displayed peptide libraries}},
url = {{http://jds.fass.org/cgi/content/abstract/87/10/3247}},
volume = {{87}},
year = {{2004}},
}