MPP notes (dja) : MTHFD1L

1. MPP OUTSIDE REQUEST FORM Reviewed 07/27/11 Requestor(s): Dean Appling Institution: U. Texas-Austin Request Title: Human mitochondrial proteins MTHFD1L and MTHFD2L

2. MPP notes (dja): MTHFD1L Function and Medical Relevance - MTHFD1L is an MPP "ListB" (Most) Biological Relevance target. See reviewer notes. Physical Characteristics-The 978 aa sequence features a transit peptide (1-31) that is cleaved to create the mature protein, an ATP-binding region (423-430), an inactive MTHFD /cyclohydrolase domain (31-348), and an FTHF synthetase domain (349-978). This is the canonical isoform; isoform 2 is much smaller (30 kda). Two lysines are acetylated. MTHFD1L is said to form a homodimer. Similar Structures-A bacterial N(10)-formyltetrahydrofolate synthetase aligning with 53% identity over aa 362-976 has been solved (1eg7a). Interactors-11 other proteins have been demonstrated to bind to MTHFD1L. Substrates are ATP, formate, tetrahydrofolate; products are ADP, phosphate , and 10-formyltetrahydrofolate. Kms for THF monoglutamate, triglutamate, and pentaglutamate have been determined. Status- MTHFD1L was selected in List B by MPP but not yet placed in a cloning workgroup. In June 2011, NESG cloned multiple variants encompassing 4 domains or fragments, approximately residues 65-350, 72-181, 182-298, and 372-968. MTHFD2L Function and Medical Relevance - See reviewer notes. Physical Characteristics-The 347 aa sequence sent by Dean Appling is 58 aa longer at the N-term than the longest isoform in Uniprot , otherwise they match. TargetP confidently predicts a transit peptide (1-8), Uniprot does not predict one. Functional domains are predicted by PFAM (1-111 and 114-288). There are two smaller alternative splicing isoforms. Similar Structures-There are 3 prokaryotic structures with 47-49% identity over most of the length of the protein, and a human folate-dependent dehydrogenase/cyclohydrolase bifunctional enzyme with 42% identity (1a4iA) with NADPH bound. Interactors-Substrates include NAD, products NADH; Mg2+ is likely to be a cofactor. Status - MPP has not yet selected this protein. NESG selected it in May 2011 and designed constructs of residues 1-112 and 113-289. Suggestions 1. Get boundaries from Dean Appling for soluble, active proteins he mentions. Pass info to NESG, and wait for results from their screens before doing any MPP work. Conclusions – suggestions accepted. Dja will contact Dean Appling, get boundaries, compare to NESG fragments, check on expression before MPP does any work.