Design, Expression, and Crystallization of Recombinant Lectin from the Garden Pea (Pisum sativum)

Prasthofer, T; Phillips, Susan R.; Suddath, F. L.; Engler, Jeffrey A.

doi:10.1016/s0021-9258(18)83499-8

Cited by 29 publications

(3 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The physiological binding partner of Psl is not known, and exploring its specificity toward monosaccharides, small oligosaccharides and modified sugars has not led to the discovery of a high affinity binding partner (11,(24)(25)(26)(27)(28). The fact, however, that Psl binds to surface oligosaccharides of red blood cells and agglutinates them (i.e., cross-links and precipitates them) has been used as a test of its activity (13,16). Unfortunately, this test had a low time resolution of about 1 day when applied to the refolding experiments, because it was necessary to concentrate the sample from the dilute refolding solutions.…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Pea Seed Lectin Folds and Oligomerizes via an Intermediate Not Represented in the Structural Hierarchy

Küster

Seckler

2008

Biochemistry

View full text Add to dashboard Cite

Large oligomeric proteins are usually thought to fold and assemble hierarchically: Domains fold and coalesce to form the subunits, and folded subunits can then associate to form the multimeric structure. We have investigated the refolding pathway of the beta-sheet protein pea seed lectin using spectroscopic and hydrodynamic techniques. In vivo, it is proteolytically processed post-translationally, so that the single-domain subunits of the initial homodimer themselves become heterodimers of intertwined fragment polypeptide chains. Despite this complex topology, mature pea seed lectin reassembles with considerable efficiency at low total protein concentration (10 mug/mL) and low temperature (10 degrees C), albeit very slowly (t1/2 approximately 2 days). Contrary to expectations, the primary assembly product is not the intact beta-sheet domain, but the larger fragment chains first dimerize to form the native-like subunit interface. The smaller fragment chains then associate with this preformed dimer.

show abstract

Section: Resultsmentioning

confidence: 99%

“…Hemagglutination Assay. The assay was carried out according to (13) and (16). In a microtiter plate with 96 round-bottomed wells, a series of 2-fold dilutions of the tested protein solution in PBS (10 mM sodium phosphate, 150 mM NaCl, pH 6.8) in a final volume of 250 µL was prepared.…”

Section: Methodsmentioning

confidence: 99%

Pea Seed Lectin Folds and Oligomerizes via an Intermediate Not Represented in the Structural Hierarchy

Küster

Seckler

2008

Biochemistry

View full text Add to dashboard Cite

show abstract

“…( 2014 ) PNA Arachis hypogaea Gal, GalNAc 1V6I Kundhavai Natchiar et al ( 2004 ) PSA Pisum sativum Glu, Man 2LTN Prasthofer et al . ( 1989 ) RPbAI Robinia pseudoacacia GalNAc 1FNY Rabijns et al . ( 2001 ) SBA Glycine max GalNAc 1SBF Olsen et al .…”

Section: Carbohydrate Specificities Of Legume Lectinsmentioning

confidence: 99%

Research advances and prospects of legume lectins

Katoch

Tripathi

2021

J Biosci

View full text Add to dashboard Cite

Effect of glycosylation on the structure of Erythrina corallodendron lectin

et al. 2004

View full text Add to dashboard Cite

The three-dimensional structure of the recombinant form of Erythrina corallodendron lectin, complexed with lactose, has been elucidated by X-ray crystallography at 2.55 A resolution. Comparison of this non-glycosylated structure with that of the native glycosylated lectin reveals that the tertiary and quaternary structures are identical in the two forms, with local changes observed at one of the glycosylation sites (Asn17). These changes take place in such a way that hydrogen bonds with the neighboring protein molecules in rECorL compensate those made by the glycan with the protein in ECorL. Contrary to an earlier report, this study demonstrates that the glycan attached to the lectin does not influence the oligomeric state of the lectin. Identical interactions between the lectin and the non-covalently bound lactose in the two forms indicate, in line with earlier reports, that glycosylation does not affect the carbohydrate specificity of the lectin. The present study, the first of its kind involving a glycosylated protein with a well-defined glycan and the corresponding deglycosylated form, provides insights into the structural aspects of protein glycosylation.

show abstract

Design, Expression, and Crystallization of Recombinant Lectin from the Garden Pea (Pisum sativum)

Cited by 29 publications

References 24 publications

Pea Seed Lectin Folds and Oligomerizes via an Intermediate Not Represented in the Structural Hierarchy

Pea Seed Lectin Folds and Oligomerizes via an Intermediate Not Represented in the Structural Hierarchy

Research advances and prospects of legume lectins

Effect of glycosylation on the structure of Erythrina corallodendron lectin

Contact Info

Product

Resources

About