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Lysine at position 222 of the goat prion protein inhibits the binding of monoclonal antibody F99/97.6.1.
MedLine Citation:
PMID:  22914824     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
Prion protein (PrP) is encoded by the PRNP gene, which is highly polymorphic in goats, with polymorphisms encoding amino acid substitutions at the protein level. In the current study, the reactivity of monoclonal antibody (mAb) F99/97.6.1 in binding PrP from goats polymorphic at PRNP codon 222 was investigated. Nervous tissue from 30 scrapie-negative goats with 3 different genotypes (222Q/Q, 222Q/K, and 222K/K) was analyzed by Western blot using mAbs P4 and F99/97.6.1. Although PrP was detected in all 30 samples by mAb P4, detection of PrP by mAb F99/97.6.1 was limited to 222Q/Q (12/12). No PrP was detected by mAb F99/97.6.1 in the 222K/K samples (n = 6), and the signal intensity of mAb F99/97.6.1 for PrP was lower for the 222Q/K samples (12/12 samples). To further investigate these results, additional Western blot analyses were performed, and the PrP signals detected by mAbs F99/97.6.1 and SAF84 were then quantified. The mean F99/SAF84 ratio (± standard deviation) calculated for the 222Q/Q group was 0.73 ± 1.26, and the mean for the 222Q/K group was 0.27 ± 1.31. Statistical analysis of these values evidenced statistically significant differences between the 222Q/Q and 222Q/K samples. The results of the study thus revealed an inhibition by lysine at position 222 on the binding of mAb F99/97.6.1 to goat PrP. This has implications for the use of mAb F99/97.6.1 for diagnostic purposes. Because the 222K allele could be a target for genetic selection in goats, the differential reactivity of mAb F99/97.6.1 could be exploited with a genotyping test setup.
Authors:
Maria Mazza; Chiara Guglielmetti; Marianna Pagano; Simona Sciuto; Francesco Ingravalle; Francesca Martucci; Maria Caramelli; Pier Luigi Acutis
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc     Volume:  24     ISSN:  1943-4936     ISO Abbreviation:  J. Vet. Diagn. Invest.     Publication Date:  2012 Sep 
Date Detail:
Created Date:  2012-08-23     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9011490     Medline TA:  J Vet Diagn Invest     Country:  United States    
Other Details:
Languages:  eng     Pagination:  971-5     Citation Subset:  IM    
Affiliation:
1Maria Mazza, Italian Reference Centre for TSEs, Istituto Zooprofilattico Sperimentale del Piemonte, Liguria e Valle d'Aosta, Via Bologna 148-10154, Turin, Italy. maria.mazza@izsto.it.
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