EurekaMag
+ Translate
+ Most Popular
The pigeon tick (Argas reflexus): its biology, ecology, and epidemiological aspects
Prevalence of hemoglobin abnormalities in Kebili (Tunisian South)
Lipogranuloma: a preventable complication of dacryocystorhinostomy
Value of basal plasma cortisol assays in the assessment of pituitary-adrenal insufficiency
Bees from the Belgian Congo. The acraensis group of Anthophora
Placing gingival retraction cord
Total serum IgE, allergy skin testing, and the radioallergosorbent test for the diagnosis of allergy in asthmatic children
Acariens plumicoles Analgesoidea parasites des oiseaux du Maroc
Injuries of terminal phalanges of the fingers in children
Biology of flowering and nectar production in pear (Pyrus communis)
Das Reliktvorkommen der Aspisviper (Vipera aspis L.) im Schwarzwald
Hydrological modelling of drained blanket peatland
Pathologic morphology and clinical significance of the anomalous origin of the left circumflex coronary artery from the right coronary artery. General review and autopsy analysis of 30 cases
Cyto genetic analyses of lymphocyte cultures after exposure to calcium cyclamate
Axelrodia riesei, a new characoid fish from Upper Rio Meta in Colombia With remarks concerning the genus Axelrodia and description of a similar, sympatric, Hyphessobrycon-species
Favorable evolution of a case of tuberculosis of pancreas under antibiotic action
RIFM fragrance ingredient safety assessment, Valencene, CAS Registry Number 4630-07-3
Parenteral microemulsions: an overview
Temperate pasture: management for grazing and conservation
Evaluation of a new coprocessed compound based on lactose and maize starch for tablet formulation
Thermal expansion and cracking of three confined water-saturated igneous rocks to 800C
Revision of the genera of the tribe Stigmoderini (Coleoptera: Buprestidae) a discussion of phylogenetic relationships
Anal tuberculosis. Report of a case
Gastric tuberculosis in the past and present
Adaptive responses of the cardiovascular system to prolonged spaceflight conditions: assessment with Holter monitoring

The interaction of Glu294 at the subunit interface is important for the activity and stability of goose delta-crystallin


The interaction of Glu294 at the subunit interface is important for the activity and stability of goose delta-crystallin



Molecular Vision 15: 2358-2363



ISSN/ISBN: 1090-0535

PMID: 19936305

delta-Crystallin is a soluble structural protein in found in avian eye lenses; it shares high amino acid sequence identity with argininosuccinate lyase. E294 is the only residue located at the double dimer interface and it performs hydrogen bonding with the active site residues of H160 and K323 in the neighboring and diagonal subunits, respectively. H160 is reported to play an important role in catalysis due to its H-bond interaction with the fumarate moiety of the substrate. In order to clarify the function of E294 in either stabilization of the quaternary structure or in catalysis, we carried out site-directed mutagenesis and functional analysis. The structure of both wild-type and mutant proteins were analyzed by circular dichroism (CD) spectroscopy, fluorescence spectra, and analytical ultracentrifugation. Structural stability was measured by CD and tryptophan fluorescence. A modeled structure of the E294L mutant was built and optimized with energy minimization. No gross structural changes were observed when E294 was substituted with leucine, as judged by circular dichroism, tryptophan fluorescence, ANS fluorescence, and sedimentation velocity analyses. However, this mutant enzyme had only about 10% of the activity of a wild-type enzyme and its secondary structure was more easily denatured by increased temperature than that of a wild-type enzyme. The mutant protein also underwent its first unfolding transition at a lower concentration of guanidinium-hydrochloride than the wild-type protein. These results indicate that the interactions offered by E294 in the dimer-dimer interface of delta-crystallin are required to maintain the hydrogen bonding network in the active site for catalysis. Disruption of the interaction had no significant effect on the conformation and quaternary structure of delta-crystallin but it did lead to instability in the double dimer structure.

Please choose payment method:






(PDF emailed within 1 workday: $29.90)

Accession: 056388117

Download citation: RISBibTeXText

Related references

Substitution of residues at the double dimer interface affects the stability and oligomerization of goose delta-crystallin. Febs Journal 276(18): 5126-5136, 2009

Critical role of tryptophanyl residues in the conformational stability of goose delta-crystallin. Experimental Eye Research 83(3): 658-666, 2006

Chaperone-like activity of alpha-crystallin. The effect of NADPH on its interaction with zeta-crystallin. Journal of Biological Chemistry 269(18): 13266-13272, 1994

Lens-preferred activity of chicken delta 1- and delta 2-crystallin enhancers in transgenic mice and evidence for retinoic acid-responsive regulation of the delta 1-crystallin gene. Developmental Genetics 20(3): 258-266, 1997

Identification of subunit-subunit interaction sites in αA-WT crystallin and mutant αA-G98R crystallin using isotope-labeled cross-linker and mass spectrometry. Plos one 8(6): E65610, 2013

Effect of phosphorylation on alpha B-crystallin: differences in stability, subunit exchange and chaperone activity of homo and mixed oligomers of alpha B-crystallin and its phosphorylation-mimicking mutant. Journal of Molecular Biology 375(4): 1040-1051, 2008

Substitution in the interface of subunit-subunit interactions enhances Escherichia coli RecA protein properties important for its recombinogenic activity. Journal of Molecular Biology 314(4): 923-935, 7 December, 2001

L29M] substitution in the interface of subunit-subunit interactions enhances Escherichia coli RecA protein properties important for its recombinogenic activity. Journal of Molecular Biology 314(4): 923-935, 2001

Stability and activity modulation of chymotrypsins in AOT reversed micelles by protein-interface interaction. Interaction of a-chymotrypsin with a negative interface leads to a cooperative breakage of a salt bridge that keeps the catalytic active conformation (Ile16-Asp194). Biotechnology and Bioengineering 59(3): 0-3, 1998

The control of delta-crystallin gene expression during lens cell development: dissociation of cell elongation, cell division, delta-crystallin synthesis, and delta-crystallin mRNA accumulation. Developmental Biology 59(2): 174-182, 1977

Mutations in human αA-crystallin/sHSP affect subunit exchange interaction with αB-crystallin. Plos one 7(2): E31421, 2012

Stability and activity modulation of chymotrypsins in AOT reversed micelles by protein-interface interaction: interaction of alpha-chymotrypsin with a negative interface leads to a cooperative breakage of a salt bridge that keeps the catalytic active conformation (Ile16-Asp194). Biotechnology and Bioengineering 59(3): 360-363, 1998

A conserved protein interaction interface on the type 5 G protein beta subunit controls proteolytic stability and activity of R7 family regulator of G protein signaling proteins. Journal of Biological Chemistry 285(52): 41100-41112, 2010

The effect of N-terminal truncation on double-dimer assembly of goose delta-crystallin. Biochemical Journal 392(Pt 3): 545-554, 2005

Facile cloning and sequence analysis of goose delta-crystallin gene based on polymerase chain reaction. Biochemical and Biophysical Research Communications 192(2): 948-953, 1993