• Identification of disordered regions in protein constructs that prevent crystallization
  
• Identification of optimum crystallization conditions
  
• Identifying the effect of point mutations on protein stability (e.g. wild-type protein vs. congenital mutations)

Proteins flex, unfold, re-fold and posses a variety of motions important to their function. Amide hydrogen/deuterium exchange, coupled with proteolysis and liquid chromatography-mass spectrometry (H/D-Ex) is becoming a popular method for probing these motions¹.  For example, large collaborative protein structure initiatives such as the NESG and JCSG regularly employ H/D-Ex to help design well folded highly stable protein constructs. This is due to the ability of H/D-Ex to reliably delineate automonous structural domains from disordered regions^2-4.

References:
1. Hamuro Y, Coales SJ, Southern MR, Nemeth-Cawley JF, Stranz DD, Griffin PR. Rapid analysis of protein structure and dynamics by hydrogen/deuterium exchange mass spectrometry. J Biomol Tech 2003;14:171-182.

2. Sharma S, Zheng H, Huang YJ, Ertekin A, Hamuro Y, Rossi P, Tejero R, Acton TB, Xiao R, Jiang M, Zhao L, Ma LC, Swapna GV, Aramini JM, Montelione GT "Construct optimization for protein NMR structure analysis using amide hydrogen/deuterium exchange mass spectrometry" Proteins. 2009 Feb 2.

3. Pantazatos D, Kim JS, Klock HE, Stevens RC, Wilson IA, Lesley SA, Woods VL, Jr. Rapid refinement of crystallographic protein construct definition employing enhanced hydrogen/deuterium exchange MS. Proc Natl Acad Sci U S A 2004;101:751-756.

4. Spraggon G, Pantazatos D, Klock HE, Wilson IA, Woods VL, Jr., Lesley SA. On the use of DXMS to produce more crystallizable proteins: structures of the T. maritima proteins TM0160 and TM1171. Protein Sci 2004;13:3187-3199.



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