6RMS
The Structure of variant D274E of the Mo-insertase domain Cnx1E from Arabidopsis thaliana in complex with AMP
Summary for 6RMS
| Entry DOI | 10.2210/pdb6rms/pdb |
| Descriptor | Molybdopterin biosynthesis protein CNX1, SULFATE ION, 1,2-ETHANEDIOL, ... (7 entities in total) |
| Functional Keywords | arabidopsis, arabidopsis proteins, metalloproteins, catalytic domain, nucleotide binding, adenosine monophosphate, plant protein |
| Biological source | Arabidopsis thaliana (Thale cress) |
| Total number of polymer chains | 1 |
| Total formula weight | 46015.07 |
| Authors | Krausze, J. (deposition date: 2019-05-07, release date: 2020-02-19, Last modification date: 2024-01-24) |
| Primary citation | Hercher, T.W.,Krausze, J.,Hoffmeister, S.,Zwerschke, D.,Lindel, T.,Blankenfeldt, W.,Mendel, R.R.,Kruse, T. Insights into the Cnx1E catalyzed MPT-AMP hydrolysis. Biosci.Rep., 40:-, 2020 Cited by PubMed Abstract: Molybdenum insertases (Mo-insertases) catalyze the final step of molybdenum cofactor (Moco) biosynthesis, an evolutionary old and highly conserved multi-step pathway. In the first step of the pathway, GTP serves as substrate for the formation of cyclic pyranopterin monophosphate, which is subsequently converted into molybdopterin (MPT) in the second pathway step. In the following synthesis steps, MPT is adenylated yielding MPT-AMP that is subsequently used as substrate for enzyme catalyzed molybdate insertion. Molybdate insertion and MPT-AMP hydrolysis are catalyzed by the Mo-insertase E-domain. Earlier work reported a highly conserved aspartate residue to be essential for Mo-insertase functionality. In this work, we confirmed the mechanistic relevance of this residue for the Arabidopsis thaliana Mo-insertase Cnx1E. We found that the conservative substitution of Cnx1E residue Asp274 by Glu (D274E) leads to an arrest of MPT-AMP hydrolysis and hence to the accumulation of MPT-AMP. We further showed that the MPT-AMP accumulation goes in hand with the accumulation of molybdate. By crystallization and structure determination of the Cnx1E variant D274E, we identified the potential reason for the missing hydrolysis activity in the disorder of the region spanning amino acids 269 to 274. We reasoned that this is caused by the inability of a glutamate in position 274 to coordinate the octahedral Mg2+-water complex in the Cnx1E active site. PubMed: 31860061DOI: 10.1042/BSR20191806 PDB entries with the same primary citation |
| Experimental method | SOLUTION SCATTERING X-RAY DIFFRACTION (1.74 Å) |
Structure validation
Download full validation report
