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1: Biochemistry. 1993 Aug 17;32(32):8146-50.Links

Dimerization of Thermomonospora fusca beta-1,4-endoglucanase E2.

McGinnis K, Kroupis C, Wilson DB.

Section of Biochemistry, Molecular, and Cell Biology, Cornell University, Ithaca, New York 14853.

Unboiled Thermomonospora fusca endoglucanase E2 electrophoresed on SDS-polyacrylamide gels migrated in the range of 80-90 kDa, but when boiled it migrated in the 40-42-kDa range. Sedimentation equilibrium centrifugation as well as chemical cross-linking experiments confirmed that E2 is a dimer. The dimer was reversibly dissociated at low pH. The E2 dimer was stable up to 70 degrees C, but began to dissociate at this temperature after a 30-60-min incubation. A nondimerizing mutant was obtained using region-specific chemical mutagenesis. DNA sequencing of this mutant revealed a single base change that substituted Gly for Glu-263. Chemical modification of carboxylic acid residues in E2 disrupted the dimer interaction.

PMID: 8347613 [PubMed - indexed for MEDLINE]