PMID- 19580466
OWN - NLM
STAT- MEDLINE
DCOM- 20100708
LR  - 20211020
IS  - 1523-4681 (Electronic)
IS  - 0884-0431 (Print)
IS  - 0884-0431 (Linking)
VI  - 25
IP  - 1
DP  - 2010 Jan
TI  - Mapping of human autoantibody binding sites on the calcium-sensing receptor.
PG  - 132-40
LID - 10.1359/jbmr.090703 [doi]
AB  - Previously, we have demonstrated the presence of anti-calcium-sensing receptor 
      (CaSR) antibodies in patients with autoimmune polyglandular syndrome type 1 
      (APS1), a disease that is characterized in part by hypoparathyroidism involving 
      hypocalcemia, hyperphosphatemia, and low serum levels of parathyroid hormone. The 
      aim of this study was to define the binding domains on the CaSR of anti-CaSR 
      antibodies found in APS1 patients and in one patient suspected of having 
      autoimmune hypocalciuric hypercalcemia (AHH). A phage-display library of CaSR 
      peptides was constructed and used in biopanning experiments with patient sera. 
      Selectively enriched IgG-binding peptides were identified by DNA sequencing, and 
      subsequently, immunoreactivity to these peptides was confirmed in ELISA. 
      Anti-CaSR antibody binding sites were mapped to amino acid residues 41-69, 
      114-126, and 171-195 at the N-terminal of the extracellular domain of the 
      receptor. The major autoepitope was localized in the 41-69 amino acid sequence of 
      the CaSR with antibody reactivity demonstrated in 12 of 12 (100%) APS1 patients 
      with anti-CaSR antibodies and in 1 AHH patient with anti-CaSR antibodies. Minor 
      epitopes were located in the 114-126 and 171-195 amino acid domains, with 
      antibody reactivity shown in 5 of 12 (42%) and 4 of 12 (33%) APS1 patients, 
      respectively. The results indicate that epitopes for anti-CaSR antibodies in the 
      AHH patient and in the APS1 patients who were studied are localized in the 
      N-terminal of the extracellular domain of the receptor. The present work has 
      demonstrated the successful use of phage-display technology in the discovery of 
      CaSR-specific epitopes targeted by human anti-CaSR antibodies.
CI  - 2010 American Society for Bone and Mineral Research
FAU - Kemp, E Helen
AU  - Kemp EH
AD  - Department of Human Metabolism, School of Medicine, University of Sheffield, 
      Sheffield S10 2JF, United Kingdom. e.h.kemp@sheffield.ac.uk
FAU - Gavalas, Nikos G
AU  - Gavalas NG
FAU - Akhtar, Samia
AU  - Akhtar S
FAU - Krohn, Kai J E
AU  - Krohn KJ
FAU - Pallais, J Carl
AU  - Pallais JC
FAU - Brown, Edward M
AU  - Brown EM
FAU - Watson, Philip F
AU  - Watson PF
FAU - Weetman, Anthony P
AU  - Weetman AP
LA  - eng
GR  - R01 DK067111/DK/NIDDK NIH HHS/United States
GR  - DK67111/DK/NIDDK NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - J Bone Miner Res
JT  - Journal of bone and mineral research : the official journal of the American 
      Society for Bone and Mineral Research
JID - 8610640
RN  - 0 (Autoantibodies)
RN  - 0 (CASR protein, human)
RN  - 0 (Epitopes)
RN  - 0 (Peptide Library)
RN  - 0 (Peptides)
RN  - 0 (Receptors, Calcium-Sensing)
SB  - IM
MH  - Adolescent
MH  - Adult
MH  - Amino Acid Sequence
MH  - Autoantibodies/*immunology
MH  - Binding Sites
MH  - Case-Control Studies
MH  - Child
MH  - Consensus Sequence
MH  - Enzyme-Linked Immunosorbent Assay
MH  - Epitopes/immunology
MH  - Female
MH  - Humans
MH  - Male
MH  - Middle Aged
MH  - Molecular Sequence Data
MH  - Peptide Library
MH  - Peptides/chemistry/immunology
MH  - Polyendocrinopathies, Autoimmune/blood/immunology
MH  - Protein Interaction Mapping/*methods
MH  - Receptors, Calcium-Sensing/*chemistry/*immunology
MH  - Young Adult
PMC - PMC3153323
EDAT- 2009/07/08 09:00
MHDA- 2010/07/09 06:00
PMCR- 2011/01/01
CRDT- 2009/07/08 09:00
PHST- 2009/07/08 09:00 [entrez]
PHST- 2009/07/08 09:00 [pubmed]
PHST- 2010/07/09 06:00 [medline]
PHST- 2011/01/01 00:00 [pmc-release]
AID - 10.1359/jbmr.090703 [doi]
PST - ppublish
SO  - J Bone Miner Res. 2010 Jan;25(1):132-40. doi: 10.1359/jbmr.090703.