• 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 function¹. Amide hydrogen/deuterium exchange, coupled with proteolysis and liquid chromatography-mass spectrometry (H/D-Ex) is becoming a popular method for probing these motions^2-4.  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^5,6.

References:
1. Huang YJ, Montelione GT. Structural biology: proteins flex to function. Nature 2005;438:36-37.

2. Engen JR, Smith DL. Investigating protein structure and dynamics by hydrogen exchange MS. Anal Chem 2001;73:256A-265A.

3. 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.

4. Hoofnagle AN, Resing KA, Goldsmith EJ, Ahn NG. Changes in protein conformational mobility upon activation of extracellular regulated protein kinase-2 as detected by hydrogen exchange. Proc Natl Acad Sci U S A 2001;98:956-961.

5. 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.

6. 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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