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MPP OUTSIDE REQUEST FORM
Review Date
07/27/11
Requestor(s): Vamsi Mootha
Institution:
Harvard
Request Title: human mitochondrial protein MTHFD2
#
Protein
Name
ORF ID
Target Request
ID
Source Org.
Target Score
pI
Residues
Size
(kDa)
1
MTHFD2
GO.111827
TR.27351
Homo sapiens
949900930190000000
8.9
350
37895
Alt.names: NMDMC
Requestor (Vamsi Mootha) notes (edited):
Bifunctional methylenetetrahydrofolate dehydrogenase/cyclohydrolase (MTHFD2) is a mitochondrial enzyme that catalyzes the conversion of
5,10-methenyltetrahydrofolate to 10-formyltetrahydrofolate, in a reaction coupled to mitochondrial NAD(+). This reaction allows for the
generation of formylmethionyl transfer RNA required for the initiation of protein synthesis in mitochondria, as well as generation of 1-carbon
units required for rapid nucleotide biosynthesis. MTHFD2 activity has been found in only embryonic or transformed cells; mitochondrial and
cytosolic paralogues, MTHFD2L and MTHFD1 respectively, both of which exhibit dinucleotide specificity for NADP(+) rather than NAD(+), perform
similar reactions in non-transformed adult tissues. Work from our laboratory has suggested that genetic silencing of MTHFD2 impairs proliferation
of transformed cells, and as such, this enzyme has emerged as a unique chemotherapeutic target. We want purified protein so we can perform
enzymatic assays on this protein which will eventually allow us to perform high-throughput assays for inhibitors. And a crystal structure may
allow us to rationally design inhibitors.
MPP notes (dja):
Function and Medical Relevance - see requestor notes
Physical Characteristics – The sequence consists of a transit peptide (1-35 predicted by both Uniprot and TargetP, 1-20 by PFAM) that is removed
in the mature protein, a tetrahydrofolate dehydrogenase/cyclohydrolase catalytic domain (38-155), and a NAD-binding domain (158-332).
MTHFD2 is predicted to form a homodimer. Lys50 is acetylated. MTHFD2 is 72% identical to its paralogue MTHFDL2.
Similar Structures - There are several structures with about 50% identity over most of the length of MTHFD2 (37-336), the closest being 1B0A,
the E. coli ortholog.
Interactors - Substrates are 5,10-methenyltetrahydrofolate and NAD+, products are 10-formyltetrahydrofolate and NADH. Mg2+ is a cofactor;
an enzyme/Mg2+ complex is required for binding of NAD+. Phosphate is also an absolute requirement for activity; it is bound by 2 Arg near the
NAD-binding site with a Km of 0.4 mM. Inhibitors are 2’,5’-ADP and NADP+. MTHFD2 binds proteins MCC, PARP3, PSMD13, and possibly
MTHFD1L.
Status - Launched 7/5/11 in MPP10 (Collaborator workgroup #3), currently in cloning. NESG has selected it. There is no other SG group
competition.