Kinetic analysis of ligand interaction with the gonococcal transferrin-iron acquisition system.
The transferrin iron acquisition system of Neisseria gonorrhoeae consists of two dissimilar transferrin binding proteins (Tbp) A and B. TbpA is a TonB dependent transporter while TbpB is a lipoprotein that makes iron acquisition from transferrin (Tf) more efficient. In an attempt to further define the individual roles of these receptors in the process of Tf-iron acquisition, the kinetics of the receptor proteins in regards to ligand association and dissociation were evaluated. Tf association with TbpB was rapid as compared to TbpA. Tf dissociation from the wild-type receptor occurred in a biphasic manner; an initial rapid release was followed by a slower dissociation over time. Both TbpA and TbpB demonstrated a two-phase release pattern; however, TbpA required both TonB and TbpB for efficient Tf dissociation from the cell surface. The roles of TbpA and TbpB in Tf dissociation were further examined, utilizing previously created HA fusion proteins. Using a Tf-utilization deficient TbpA-HA mutant, we concluded that the slower rate of ligand dissociation demonstrated by the wild-type transporter was a function of successful iron internalization. Insertion into the C-terminus of TbpB decreased the rate of Tf dissociation, while insertion into the N-terminus had no effect on this process. From these studies, we propose that TbpA and TbpB function synergistically during the process of Tf iron acquisition and that TbpB makes the process of Tf-iron acquisition more efficient at least in part by affecting association and dissociation of Tf from the cell surface.
DeRocco
Amanda J
AJ
Department of Microbiology, Virginia Commonwealth University Medical Center, PO Box 980678, Richmond, VA 23298-0678, USA.
Yost-Daljev
Mary Kate
MK
Kenney
Christopher D
CD
Cornelissen
Cynthia Nau
CN
D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D007501:Iron / Q000378:metabolism*; D007700:Kinetics; D009344:Neisseria gonorrhoeae / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding / Q000502:physiology; D014168:Transferrin / Q000235:genetics / Q000378:metabolism*; D033881:Transferrin-Binding Protein A / Q000235:genetics / Q000378:metabolism*; D033882:Transferrin-Binding Protein B / Q000235:genetics / Q000378:metabolism*
D005656:Fungal Proteins; D014168:Transferrin; D033881:Transferrin-Binding Protein A; D033882:Transferrin-Binding Protein B; D007501:Iron