Specific glycosidase activity isolated from a random phage display antibody library

G. N. Goud, O. Artsaenko, M. Bols, Michael Sierks

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Carbohydrates serve as key receptor sites in various cellular events such as viral attachment, tumor formation, and tissue inflammation. A potential route to control these events is to manipulate targeted carbohydrate structures in vivo using specifically designed glycohydrolases. Here we show that a stereospecific catalytic activity designed toward a particular sugar and linkage can be readily isolated from a phage display antibody library derived from a nonimmunized host. The activity was isolated using a transition-state analogue mimicking an α-glucosidasic linkage as antigen and showed a 20-fold specificity for that sugar and linkage. The DNA sequence, however, contains a large deletion in the antibody gene, which also changes the downstream reading frame, resulting in a translated sequence containing only 57 amino acids that has a predominantly hydrophobic amino terminal and a strongly hydrophilic carboxy terminal. The isolated catalytic activity has a strong Ph dependence, attributable to one or more of the numerous potentially charged groups in the carboxyl terminal. While the protein readily forms more stable multimers, the 7.3-Kd monomer represents by far the smallest glycosidase enzyme reported to date and can provide substantial new information toward understanding and modifying glycosidase activity.

Original languageEnglish (US)
Pages (from-to)197-202
Number of pages6
JournalBiotechnology Progress
Volume17
Issue number1
DOIs
StatePublished - 2001
Externally publishedYes

Fingerprint

glycosidases
Glycoside Hydrolases
bacteriophages
Bacteriophages
linkage (genetics)
catalytic activity
antibodies
Antibodies
Carbohydrates
carbohydrate structure
sugars
Reading Frames
inflammation
Inflammation
antigens
carbohydrates
Antigens
nucleotide sequences
Amino Acids
receptors

ASJC Scopus subject areas

  • Food Science
  • Biotechnology
  • Microbiology

Cite this

Specific glycosidase activity isolated from a random phage display antibody library. / Goud, G. N.; Artsaenko, O.; Bols, M.; Sierks, Michael.

In: Biotechnology Progress, Vol. 17, No. 1, 2001, p. 197-202.

Research output: Contribution to journalArticle

Goud, G. N. ; Artsaenko, O. ; Bols, M. ; Sierks, Michael. / Specific glycosidase activity isolated from a random phage display antibody library. In: Biotechnology Progress. 2001 ; Vol. 17, No. 1. pp. 197-202.
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