Identification of transglutaminase substrates in HT29 colon cancer cells: use of 5-(biotinamido)pentylamine as a transglutaminase-specific probe.

Abstract

A biotinamine probe, 5-(biotinamido)pentylamine, was used for biotin-labeling of proteins in HT29 colon cancer cell extracts by endogenous transglutaminase activity. The biotin-labeled protein substrates were isolated and recovered by avidin-affinity chromatography. The proteins were separated using SDS-polyacrylamide gel electrophoresis, electroblotted onto a polyvinylidene difluoride membrane, visualized using Coomassie blue, cut out, and sequenced. Amino acid sequence data identified human fructose-1,6-bisphosphate aldolase A, an intracellular protein, as a substrate for cellular transglutaminase.

• Authors: K N Lee, M D Maxwell, M K Patterson Jr, P J Birckbichler, E Conway
• Journal: Biochimica et biophysica acta (Biochim Biophys Acta) Vol. 1136 Issue 1 Pg. 12-6 (Jul 22 1992) ISSN: 0006-3002 [Print] Netherlands
• PMID: 1353685 (Publication Type: Journal Article)

Chemical References

• Amines
• Molecular Probes
• 5-(biotinamido)pentylamine
• Biotin
• Transglutaminases
• Fructose-Bisphosphate Aldolase

Topics

• Amines
• Amino Acid Sequence
• Biotin (analogs & derivatives)
• Blotting, Western
• Chromatography, Affinity
• Colonic Neoplasms
• Fructose-Bisphosphate Aldolase (isolation & purification, metabolism)
• Humans
• Molecular Probes
• Molecular Sequence Data
• Substrate Specificity
• Transglutaminases (metabolism)
• Tumor Cells, Cultured