Probing molecular interaction between concanavalin A and mannose ligands by means of SFM.
Recently, the scanning force microscope (SFM) has been widely used for direct monitoring of specific interactions between biologically active molecules. Such studies have employed the SFM liquid-cell setup, which allows measurements to be made in the native environment with force resolution down to a tenth of a picoNewton. In this study, the ligand-receptor strength of monoclonal anti-human prostatic acid phosphatase and prostatic acid phosphatase, representing an antigen-antibody system with a single type of interaction, was determined. Then, the interaction force occurring between concanavalin A and the carbohydrate component of the glycoproteins arylsulfatase A and carboxypeptidase Y was measured. High mannose-type glycans were sought on the human prostate carcinoma cell surface. Application of an analysis based on the Poisson distribution of the number of bonds formed in all these measured systems allowed the strength of the molecular interaction to be calculated. The values of the force acting between two single molecules were 530+/-25, 790+/-32, and 940+/-39 pN between prostatic acid phosphatase and monoclonal anti-human prostatic acid phosphatase, between concanavalin A and arylsulfatase A, and between concanavalin A and carboxypeptidase Y, respectively. The value calculated from data collected for the force between concanavalin A and mannose-containing ligands present on the surface of human prostate carcinoma cells was smaller, 116+/-17 pN. The different values of the binding force between concanavalin A and mannose-containing ligands were attributed to the structural changes of the carbohydrate components.
Lekka
M
M
The Henryk Niewodniczański Institute of Nuclear Physics, Radzikowskiego 152, 31-342 Cracow, Poland. Malgorzata.Lekka@ifj.edu.pl
Laidler
P
P
Dulińska
J
J
Łabedź
M
M
Pyka
G
G
D001665:Binding Sites; D002448:Cell Adhesion / Q000502:physiology*; D045744:Cell Line, Tumor; D003208:Concanavalin A / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure*; D006801:Humans; D008024:Ligands; D008297:Male; D008358:Mannose / Q000378:metabolism*; D008846:Micromanipulation / Q000379:methods; D018625:Microscopy, Atomic Force / Q000379:methods*; D010812:Physical Stimulation / Q000379:methods; D011471:Prostatic Neoplasms / Q000503:physiopathology* / Q000648:ultrastructure*; D011485:Protein Binding; D011487:Protein Conformation; D013314:Stress, Mechanical
D008024:Ligands; D003208:Concanavalin A; D008358:Mannose