Molecular investigation of the functional relevance of missense variants of ICAM-1

In genome-wide studies, the intercellular adhesion molecule-1 (ICAM-1) locus has been associated with cardiovascular and inflammatory bowel diseases. To determine the functional relevance of five missense ICAM-1 variants (G241R; I316V; P352L; K469E; R478W), we generated wild-type and variant proteins [M2(241 R); M3(469E); M4(352L); M5(478W); M6(316V); M7(352L/ 469E)] and transiently transfected CV1 cells. Reverse transcription PCR, western blot, and ELISA did not reveal any differences in mRNA and protein expression levels for any construct. Conversely, in pulse-chase experiments, compared with wild-type (90-120 min), M3 and M5 possessed a prolonged half-life of similar to 150 min, whereas M2, M4, and M7 displayed a decreased half-life of similar to 60-75 min, implying differences in protein degradation. Our results do not indicate a major impact of missense variants on ICAM-1 biological function, even if G241R and K469E were functional in pulse-chase experiments. Whether these differences in protein stability exert measurable functional consequences needs to be elucidated further. Pharmacogenetics and Genomics 18:1017-1019 (C) 2008 Wolters Kluwer Health vertical bar Lippincott Williams & Wilkins.