A method for the general identification of protein crystals in crystallization experiments using a noncovalent fluorescent dye.
A technique is described whereby the addition of low concentrations (millimolar to micromolar) of the fluorescent dye 1,8-ANS to the protein solution prior to crystallization results in crystallization experiments in which protein crystals are strongly contrasted above background artifacts when exposed to low-intensity UV radiation. As 1,8-ANS does not covalently modify the protein sample, no further handling or purification steps are necessary. The system has been tested on a wide variety of protein samples and it has been shown that the addition of 1,8-ANS has no discernible effect on the crystallization frequencies or crystallization conditions of these proteins. As 1,8-ANS interacts with a wide variety of proteins, this is proposed to be a general solution for the automated classification of protein crystallization images and the detection of protein crystals. The results also demonstrate the expected discrimination between salt and protein crystals, as well as allowing the straightforward identification of small crystals that grow in precipitate or under a protein skin.
Groves
Matthew R
MR
EMBL Outstation Hamburg, c/o DESY, Building 25a, Notkestrasse 85, 22603 Hamburg, Germany. groves@embl-hamburg.de
Müller
Ingrid B
IB
Kreplin
Xandra
X
Müller-Dieckmann
Jochen
J
D000817:Anilino Naphthalenesulfonates / Q000737:chemistry*; D003460:Crystallization; D018360:Crystallography, X-Ray; D043364:Endo-1,4-beta Xylanases / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry*; D007328:Insulin / Q000737:chemistry; D009113:Muramidase / Q000737:chemistry; D010447:Peptide Hydrolases / Q000737:chemistry; D011506:Proteins / Q000737:chemistry*; D014357:Trypsin / Q000737:chemistry
D000817:Anilino Naphthalenesulfonates; D005456:Fluorescent Dyes; D007328:Insulin; D011506:Proteins; C027132:1-anilino-8-naphthalenesulfonate; D009113:Muramidase; D043364:Endo-1,4-beta Xylanases; D010447:Peptide Hydrolases; D014357:Trypsin