Isoelectric focusing studies of concanavalin A and the lentil lectin

Lokesh Bhattacharyya, Curtis F. Brewer

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Isoelectric focusing (IEF) of metallized and demetallized preparations of concanavalin A (Con A) consisting of either intact or fragmented subunits shows different band patterns. Metallized Con A consisting of intact polypeptide chains (intact Con A) has an isoelectric point (pI) 8.35. Metallized preparations consisting of fragmented chains (fragmented Con A) show three bands with pI values 8.0, 7.8 and 7.7. Demetallized intact Con A (intact apoCon A) has a pI of 6.5, however, it undergoes pH dependent association during IEF under certain conditions, which gives rise to multiple bands. Ampholyte-mediated demetallization of intact and fragmented Con A and subsequent aggregation of the apoprotein results in multiple bands during IEF in the presence of the pH range 3 to 10 ampholytes. However, ampholytes of the pH range 7 to 9 do not demetallize the proteins and show a single band with intact Con A. The pI of intact Con A remains essentially the same in the presence of inhibitory sugar. Furthermore, differnt moleculars forms of Con A, including locked and unlocked conformers of intact apoCon A, and the dimeric and tetramic states of both intact Con A and intact apoCon A have been identified and their pI values determined. IEF of the lentil isoelectins, LcH-A and LcH-B, shows single bands of pI 8.5 and 9.0, respectively. However, the native lectin mixture gives rise to an additional band of pI 8.8 due to a hybrid protein formed by ampholyte-mediated subunit exchange between the isolectins.

Original languageEnglish (US)
Pages (from-to)131-142
Number of pages12
JournalJournal of Chromatography A
Volume502
Issue numberC
DOIs
StatePublished - 1990

Fingerprint

Isoelectric Focusing
Concanavalin A
Buffers
Lectins
Lens Plant
lentil lectin
Apoproteins
Isoelectric Point
Sugars
Proteins
Agglomeration
Peptides

ASJC Scopus subject areas

  • Analytical Chemistry
  • Clinical Biochemistry
  • Molecular Medicine

Cite this

Isoelectric focusing studies of concanavalin A and the lentil lectin. / Bhattacharyya, Lokesh; Brewer, Curtis F.

In: Journal of Chromatography A, Vol. 502, No. C, 1990, p. 131-142.

Research output: Contribution to journalArticle

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