Effect of vitamin B6 availability on serine hydroxymethyltransferase in MCF-7 cells.
Folate-activated one-carbon units are derived from serine through the activity of the pyridoxal-phosphate (PLP)-dependent isozymes of serine hydroxymethyltransferase (SHMT). The effect of vitamin B(6) availability on the activity and expression of the human mitochondrial and cytoplasmic SHMT isozymes was investigated in human MCF-7 cells. Cells were cultured for 6 months in vitamin B(6) replete (4.9 microM pyridoxine) minimal essential medium (alphaMEM) or vitamin B(6)-deficient medium containing 49, 4.9 or 0.49 nM pyridoxine. Total cellular PLP levels and SHMT activity were reduced 72% and 7%, respectively, when medium pyridoxine was decreased from 4.9 microM to 49 nM. Cells cultured in medium containing 4.9 nM pyridoxine exhibited 75%, 27% and 60% reduced levels of PLP, SHMT activity and S-adenosylmethionine, respectively, compared to cells cultured in alphaMEM. Cytoplasmic SHMT activity and protein levels, but not mRNA levels, were decreased in cells cultured in vitamin B(6) deficient medium, whereas mitochondrial SHMT activity and protein levels were less sensitive to vitamin B(6) availability. PLP bound to cytoplasmic SHMT with a K(d)=850 nM, a value two orders of magnitude lower than previously reported for the bovine cytoplasmic SHMT isozyme. Collectively, these data indicate that vitamin B(6) restriction decreases the activity and stability of SHMT, and that the cytoplasmic isozyme is more sensitive to vitamin B(6) deficiency than the mitochondrial isozyme in MCF-7 cells.
Perry
Cheryll
C
Cornell University, Division of Nutritional Science, Ithaca, NY 14853, USA.
Yu
Sun
S
Chen
Jaclyn
J
Matharu
Kabir S
KS
Stover
Patrick J
PJ
D045744:Cell Line, Tumor; D003593:Cytoplasm / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005492:Folic Acid / Q000737:chemistry; D015971:Gene Expression Regulation, Enzymologic; D015972:Gene Expression Regulation, Neoplastic; D012696:Glycine Hydroxymethyltransferase / Q000378:metabolism*; D006710:Homocysteine / Q000737:chemistry; D006801:Humans; D007700:Kinetics; D008928:Mitochondria / Q000378:metabolism; D020033:Protein Isoforms; D011736:Pyridoxine / Q000737:chemistry; D012436:S-Adenosylmethionine / Q000378:metabolism; D025101:Vitamin B 6 / Q000378:metabolism*
D020033:Protein Isoforms; D006710:Homocysteine; D012436:S-Adenosylmethionine; D025101:Vitamin B 6; D005492:Folic Acid; D012696:Glycine Hydroxymethyltransferase; D011736:Pyridoxine