PMID- 19179454
OWN - NLM
STAT- MEDLINE
DCOM- 20090622
LR  - 20100322
IS  - 1479-6813 (Electronic)
IS  - 0952-5041 (Linking)
VI  - 42
IP  - 4
DP  - 2009 Apr
TI  - Calcium-sensing receptor mutations and denaturing high performance liquid 
      chromatography.
PG  - 331-9
LID - 10.1677/JME-08-0164 [doi]
AB  - The calcium-sensing receptor (CASR), a plasma membrane G-protein-coupled 
      receptor, is expressed in parathyroid gland and kidney, and controls systemic 
      calcium homeostasis. Inactivating CASR mutations are associated with familial 
      hypocalciuric hypercalcemia (FHH) and neonatal severe hyperparathyroidism, and 
      activating mutations cause autosomal dominant hypocalcemia (ADH). CASR mutation 
      identification plays an important role in the clinical management of mineral 
      metabolism disorders. We describe here a high-throughput method using screening 
      with denaturing high performance liquid chromatography (DHPLC) to initially 
      interrogate 12 amplicons covering translated exons and exon/intron boundaries, 
      followed by sequencing of any amplicon with a modified melting curve relative to 
      wild type, and direct sequencing of a 13th amplicon encoding the COOH-terminal 
      tail to distinguish causative mutations from three common missense single 
      nucleotide polymorphisms. A blinded analysis of 32 positive controls representing 
      mutations throughout the CASR sequence, as well as 22 negative controls, yielded 
      a concordance rate of 100%. We report eight novel and five recurrent FHH 
      mutations, along with six novel and two recurrent ADH mutations. Thus, DHPLC 
      provides a rapid and effective means to screen for CASR mutations.
FAU - Cole, David E C
AU  - Cole DE
AD  - Departments of Laboratory Medicine and Pathobiology, Medicine, and Genetics, 
      University of Toronto, Ontario, Canada.
FAU - Yun, Francisco H J
AU  - Yun FH
FAU - Wong, Betty Y L
AU  - Wong BY
FAU - Shuen, Andrew Y
AU  - Shuen AY
FAU - Booth, Ronald A
AU  - Booth RA
FAU - Scillitani, Alfredo
AU  - Scillitani A
FAU - Pidasheva, Svetlana
AU  - Pidasheva S
FAU - Zhou, Xiang
AU  - Zhou X
FAU - Canaff, Lucie
AU  - Canaff L
FAU - Hendy, Geoffrey N
AU  - Hendy GN
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20090129
PL  - England
TA  - J Mol Endocrinol
JT  - Journal of molecular endocrinology
JID - 8902617
RN  - 0 (Mutant Proteins)
RN  - 0 (Receptors, Calcium-Sensing)
SB  - IM
EIN - J Mol Endocrinol. 2010 Apr;44(4):257
MH  - Chromatography, High Pressure Liquid/*methods
MH  - Computational Biology
MH  - DNA Mutational Analysis
MH  - Exons/genetics
MH  - Genome, Human/genetics
MH  - Humans
MH  - Mutant Proteins/genetics
MH  - Mutation/*genetics
MH  - Mutation, Missense/genetics
MH  - Receptors, Calcium-Sensing/*genetics
MH  - Reproducibility of Results
EDAT- 2009/01/31 09:00
MHDA- 2009/06/23 09:00
CRDT- 2009/01/31 09:00
PHST- 2009/01/31 09:00 [entrez]
PHST- 2009/01/31 09:00 [pubmed]
PHST- 2009/06/23 09:00 [medline]
AID - JME-08-0164 [pii]
AID - 10.1677/JME-08-0164 [doi]
PST - ppublish
SO  - J Mol Endocrinol. 2009 Apr;42(4):331-9. doi: 10.1677/JME-08-0164. Epub 2009 Jan 
      29.