by Peter Nollert
June 16, 2012 01:06
If only we would not have to ‘waste’ so much protein sample for mind-numbing trial and error crystallization experiments. Aside from clever pre-crystallization screening approaches, there are now several reports that indicate that we may start to get a handle to more rational approaches. This is the first short review of such a report towards shortening the path from protein sample to crystals.
The observation is that ‘Immobilized metal-affinity chromatography protein-recovery screening is predictive of crystallographic structure success’. A post-mortem analysis of purification and crystallization data has revealed quantitatively what many of us have picked up at the bench: proteins that purify well are easy to crystallize.
Choi R, Kelley A, Leibly D, Hewitt SN, Napuli A, & Van Voorhis W (2011). Immobilized metal-affinity chromatography protein-recovery screening is predictive of crystallographic structure success. Acta crystallographica. Section F, Structural biology and crystallization communications, 67 (Pt 9), 998-1005 PMID: 21904040
The authors of this paper however have taken this observation as a starting point and have devised a low cost IMAC high-throughput protocol (using multichannel pipettors, affinity beads and filter plates; the procedure is described in exquisite detail; thank you very much!). They applied this protocol to more than 4330 proteins (SSGCID effort) to mine IMAC recovery data for rules and find that they determined more than twice as many structures of those proteins that showed a high IMAC recovery, as compared to those with a low IMAC recovery.
Factor 2.
Not bad.