Enhancing protein crystallization through precipitant synergy.
Suitable conditions for protein crystallization are commonly identified by screening combinations of independent factors that affect crystal formation. Because precipitating agents are prime determinants of crystallization, we investigated whether a systematic exploration of combinations of mechanistically distinct precipitants would enhance crystallization. A crystallization screen containing 64 precipitant mixtures was devised. Tests with ten HIV envelope-related proteins demonstrated that use of precipitant mixtures significantly enhanced both the probability of crystallization as well as the quality of optimized crystals. Tests with hen egg white lysozyme generated a novel C2 crystal from a salt/organic solvent mixture; structure solution at 2 A resolution revealed a lattice held together by both hydrophobic and electrostatic dyad interactions. The results indicate that mechanistically distinct precipitants can synergize, with precipitant combinations adding unique dimensions to protein crystallization.
Majeed
Shahzad
S
Vaccine Research Center, National Institutes of Health, Bethesda, MD 20892, USA.
Ofek
Gilad
G
Belachew
Adam
A
Huang
Chih-chin
CC
Zhou
Tongqing
T
Kwong
Peter D
PD
D000818:Animals; D011232:Chemical Precipitation; D002645:Chickens; D003460:Crystallization; D018360:Crystallography, X-Ray; D008958:Models, Molecular; D009113:Muramidase / Q000737:chemistry; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry*
D011506:Proteins; D009113:Muramidase