New approach for the identification of folate-related pathways in human embryogenesis.
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.
Bliek
J B
JB
Departments of Obstetrics and Gynecology, Erasmus MC University Medical Center, 3000 CA Rotterdam, The Netherlands.
de Klein
A
A
Luider
Th M
TM
Lindemans
J
J
Hulsman
L
L
Guzel
C
C
de Groot
C J M
CJ
Steegers-Theunissen
R P M
RP
D001402:B-Lymphocytes / Q000378:metabolism; D002461:Cell Line, Transformed; D045744:Cell Line, Tumor; D019992:Databases as Topic; D030562:Databases, Protein; D004622:Embryo, Mammalian / Q000378:metabolism*; D004626:Embryology / Q000379:methods; D005492:Folic Acid / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D004854:Herpesvirus 4, Human / Q000378:metabolism; D006801:Humans; D008214:Lymphocytes / Q000378:metabolism; D013058:Mass Spectrometry; D010455:Peptides / Q000737:chemistry; D040901:Proteomics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013763:Tetrahydrofolates / Q000494:pharmacology; D013997:Time Factors; D014357:Trypsin / Q000494:pharmacology
D010455:Peptides; D013763:Tetrahydrofolates; D005492:Folic Acid; D014357:Trypsin; C005984:5-methyltetrahydrofolate