%0 Journal Article
%T Discrimination of three mutational events that result in a disruption of the R122 primary autolysis site of the human cationic trypsinogen (PRSS1) by denaturing high performance liquid chromatography
%A Cedric Le Mar¨¦chal
%A Jian-Min Chen
%A Isabelle Qu¨¦r¨¦
%A Odile Ragu¨¦n¨¨s
%A Claude F¨¦rec
%A Jean Auroux
%J BMC Genetics
%D 2001
%I BioMed Central
%R 10.1186/1471-2156-2-19
%X DNA samples containing either the known c.365G>A or c.365~366GC>AT variant in exon 3 of PRSS1 were used as positive controls to establish a denaturing high performance liquid chromatography (DHPLC) assay.DHPLC could readily discriminate the two known different mutational events resulting in the R122H mutation. More importantly, under the same experimental conditions, it identified a further mutational event that also occurs in the R122 primary autolysis site but results in a different amino acid substitution: c.364C>T (CGC>TGC; R122C).A rapid, simple, and low-cost assay for detecting both the known and new mutations occuring in the R122 primary autolysis site of PRSS1 was established. In addition, the newly found R122C variant represents a likely pancreatitis-predisposing mutation.Trypsin plays a central role in pancreatic exocrine physiology by acting as the trigger enzyme that leads to the activation of all the digestive proenzymes. To prevent premature trypsin activation within the pancreas, the body has evolved a series of defensive mechanisms. Of particular significance is the "built-in" R122 primary autolysis site of mammalian trypsinogens, which serves as a "self-destruct" or "fail-safe" mechanism to prevent pancreatic autodigestion [1]. Disruption of this site by a missense mutation ¨C R122H (originally termed R117H in the chymotrypsin numbering system [2]; for a discussion of mutation nomenclature see [3,4]) ¨C in the human cationic trypsinogen (PRSS1; OMIM 276000) has been associated with most of the large kindreds with hereditary pancreatitis (HP; OMIM 167800) [1].The initially identified R122H mutation results from a c.365G>A (CGC>CAC) transition in exon 3 of PRSS1[2]. Since this mutation creates a novel restriction recognition site for Alf III (A¡ýCRYGT), a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay was established [2] and has been widely apdopted to detect this most frequent mutation in both hereditary and sporadic
%U http://www.biomedcentral.com/1471-2156/2/19