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
<p>Abstract</p> <p>Background</p> <p>R122, the primary autolysis site of the human cationic trypsinogen (PRSS1), constitutes an important "self-destruct" or "fail-safe" defensive mechanism against premature trypsin activation within the pancreas. Dis...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2001-11-01
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| Series: | BMC Genetics |
| Online Access: | http://www.biomedcentral.com/1471-2156/2/19 |
| _version_ | 1811302725659394048 |
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| author | Férec Claude Raguénès Odile Quéré Isabelle Chen Jian-Min Le Maréchal Cedric Auroux Jean |
| author_facet | Férec Claude Raguénès Odile Quéré Isabelle Chen Jian-Min Le Maréchal Cedric Auroux Jean |
| author_sort | Férec Claude |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>R122, the primary autolysis site of the human cationic trypsinogen (PRSS1), constitutes an important "self-destruct" or "fail-safe" defensive mechanism against premature trypsin activation within the pancreas. Disruption of this site by a missense mutation, R122H, was found to cause hereditary pancreatitis. In addition to a c.365G>A (C<b>G</b>C>C<b>A</b>C) single nucleotide substitution, a c.365~366GC>AT (C<b>GC</b>>C<b>AT</b>) gene conversion event in exon 3 of <it>PRSS1</it> was also found to result in a R122H mutation. This imposes a serious concern on the genotyping of pancreatitis by a widely used polymerase chain reaction-restriction fragment length polymorphism assay, which could only detect the commonest c.365G>A variant.</p> <p>Materials and methods</p> <p>DNA samples containing either the known c.365G>A or c.365~366GC>AT variant in exon 3 of <it>PRSS1</it> were used as positive controls to establish a denaturing high performance liquid chromatography (DHPLC) assay.</p> <p>Results</p> <p>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 (<b>C</b>GC><b>T</b>GC; R122C).</p> <p>Conclusions</p> <p>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.</p> |
| first_indexed | 2024-04-13T07:34:50Z |
| format | Article |
| id | doaj.art-2bcb2ce8b65646a68f48dc2ace9b7291 |
| institution | Directory Open Access Journal |
| issn | 1471-2156 |
| language | English |
| last_indexed | 2024-04-13T07:34:50Z |
| publishDate | 2001-11-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Genetics |
| spelling | doaj.art-2bcb2ce8b65646a68f48dc2ace9b72912022-12-22T02:56:13ZengBMCBMC Genetics1471-21562001-11-01211910.1186/1471-2156-2-19Discrimination 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 chromatographyFérec ClaudeRaguénès OdileQuéré IsabelleChen Jian-MinLe Maréchal CedricAuroux Jean<p>Abstract</p> <p>Background</p> <p>R122, the primary autolysis site of the human cationic trypsinogen (PRSS1), constitutes an important "self-destruct" or "fail-safe" defensive mechanism against premature trypsin activation within the pancreas. Disruption of this site by a missense mutation, R122H, was found to cause hereditary pancreatitis. In addition to a c.365G>A (C<b>G</b>C>C<b>A</b>C) single nucleotide substitution, a c.365~366GC>AT (C<b>GC</b>>C<b>AT</b>) gene conversion event in exon 3 of <it>PRSS1</it> was also found to result in a R122H mutation. This imposes a serious concern on the genotyping of pancreatitis by a widely used polymerase chain reaction-restriction fragment length polymorphism assay, which could only detect the commonest c.365G>A variant.</p> <p>Materials and methods</p> <p>DNA samples containing either the known c.365G>A or c.365~366GC>AT variant in exon 3 of <it>PRSS1</it> were used as positive controls to establish a denaturing high performance liquid chromatography (DHPLC) assay.</p> <p>Results</p> <p>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 (<b>C</b>GC><b>T</b>GC; R122C).</p> <p>Conclusions</p> <p>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.</p>http://www.biomedcentral.com/1471-2156/2/19 |
| spellingShingle | Férec Claude Raguénès Odile Quéré Isabelle Chen Jian-Min Le Maréchal Cedric Auroux Jean 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 BMC Genetics |
| title | 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 |
| title_full | 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 |
| title_fullStr | 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 |
| title_full_unstemmed | 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 |
| title_short | 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 |
| title_sort | 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 |
| url | http://www.biomedcentral.com/1471-2156/2/19 |
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