New approach for the identification of folate-related pathways in human embryogenesis

• Published in: Cellular and molecular biology (Noisy-le-Grand, France) Vol. 50; no. 8; pp. 939 - 944
• Main Authors: Bliek, J B, de Klein, A, Luider, Th M, Lindemans, J, Hulsman, L, Guzel, C, de Groot, C J M, Steegers-Theunissen, R P M
• Format: Journal Article
• Language: English
• Published: France 01-12-2004
• Subjects: B-Lymphocytes - metabolism; Cell Line, Transformed; Cell Line, Tumor; Databases as Topic; Databases, Protein; Embryo, Mammalian - metabolism; Embryology - methods; Folic Acid - metabolism; Gene Expression Regulation, Developmental; Herpesvirus 4, Human - metabolism; Humans; Lymphocytes - metabolism; Mass Spectrometry; Peptides - chemistry; Proteomics; Reverse Transcriptase Polymerase Chain Reaction; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Tetrahydrofolates - pharmacology; Time Factors; Trypsin - pharmacology
• ISSN: 0145-5680

The role of natural folate intake and synthetic folic acid supplementation in the prevention of some congenital malformations is known, but on a molecular biological level poorly understood. In a first approach to identify folate-regulated pathways in human embryogenesis, tryptic digests of Epstein Barr Virus-immortalized B-lymphoblasts proteins from 6 cleft lip and/or palate patients and 2 controls were compared using matrix assisted laser desorption ionisation--time of flight (MALDI-TOF) mass spectrometry. After immortalisation, the lymphoblasts were cultured for 22 days in folate-rich, i.e. 5-methyltetrahydrofolate (5-mTHF), or folate-free medium. On day 22, 5-mTHF was added to the folate-free cultures and the profiles on day 22 and 23 were compared. After background correction for the peptide profiles of the folate-rich cultures, we found in the folate-free mediaseveral differentially expressed peptide peaks upon addition of 5-mTHF. These peptide peaks were mass annotated and matched withthe MSDB human database. The results suggest some folate-regulated protein candidates as Frizzled and the Rho GTP-ases WRCH and Chp that are known in human embryogenesis. Differential folate expressed proteins in patients and controls, however, have to be further investigated.