Photoaffinity labeling of a bacterial sialidase with an aryl azide derivative of sialic acid

Journal Article · Thu Jul 05 04:00:00 EDT 1990 · Journal of Biological Chemistry; (USA)

OSTI ID:6874040

van der Horst, G T; Mancini, G M; Brossmer, R; Rose, U; Verheijen, F W ^[1]

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1. Erasmus Univ., Rotterdam (Netherlands)

A photoreactive radioiodinatable derivative of 2-deoxy-2,3-didehydro-5-N-acetylneuraminic acid (NeuAc2en), 5-N-acetyl-9-(4-azidosalicoylamido)-2-deoxy-2,3-didehydroneuram inic acid (ASA-NeuAc2-en) has been synthesized and used to label the active site of Clostridium perfringens sialidase. Like NeuAc2en, its aryl azide derivative is a strong competitive inhibitor of sialidase (Ki approximately 15 microM). The absorbance spectrum of ASA-NeuAc2en shows a characteristic aryl azide peak, which disappears upon photolysis with UV light. When its radioiodinated counterpart 5-N-acetyl-9-(4-iodoazidosalicoylamido)-2-deoxy-2,3-didehydrone uraminic acid (({sup 125}I)IASA-NeuAc2en) was photolyzed in the presence of C. perfringens sialidase a 72-kDa protein was labeled. Labeling occurred specifically in the active site since it was inhibited in the presence of NeuAc2en. Chemical cleavage of the photoaffinity-labeled 72-kDa protein demonstrates that specifically labeled peptides involved in the formation of the active site can easily be determined. ASA-NeuAc2en is a valuable new tool for the identification and structural/functional analysis of sialidases and other proteins, recognizing this sialic acid derivative.

• OSTI ID: 6874040
• Journal Information: Journal of Biological Chemistry; (USA), Journal Name: Journal of Biological Chemistry; (USA) Vol. 265:19; ISSN 0021-9258; ISSN JBCHA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
560120 -- Radiation Effects on Biochemicals
Cells
& Tissue Culture
59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ABSORPTION SPECTRA
AFFINITY
BACTERIA
BETA DECAY RADIOISOTOPES
CARBOHYDRATES
CHEMICAL RADIATION EFFECTS
CHEMISTRY
CLOSTRIDIUM
DAYS LIVING RADIOISOTOPES
ELECTROMAGNETIC RADIATION
ELECTRON CAPTURE RADIOISOTOPES
ELECTROPHORESIS
ENZYME INHIBITORS
ENZYMES
GLYCOSYL HYDROLASES
HYDROLASES
INTERMEDIATE MASS NUCLEI
IODINE 125
IODINE ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
LABELLING
MICROORGANISMS
MOLECULAR STRUCTURE
MOLECULAR WEIGHT
MONOSACCHARIDES
NUCLEI
O-GLYCOSYL HYDROLASES
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
PEPTIDES
PHOTOCHEMISTRY
PROTEINS
RADIATION CHEMISTRY
RADIATION EFFECTS
RADIATIONS
RADIOISOTOPES
SACCHARIDES
SIALIC ACID
SPECTRA
SPECTROPHOTOMETRY
TRACER TECHNIQUES
ULTRAVIOLET RADIATION