Title Information
Title
Biogenesis of Trypanosome Lytic Factor and Mechanism of Trypanosome Killing
Name: Personal
Name Part
Widener, Justin W.
Role
Role Term: Text
creator
Origin Information
Copyright Date (keyDate="yes", encoding="w3cdtf")
2008
Physical Description
Extent
xx, 79 p.
digitalOrigin
born digital
Note
Thesis (Ph.D.) -- Brown University (2008)
Name: Personal
Name Part
Hajduk, Stephen
Role
Role Term: Text
director
Name: Personal
Name Part
Bennet, Richard
Role
Role Term: Text
reader
Name: Personal
Name Part
Campbell, Andrew
Role
Role Term: Text
reader
Name: Personal
Name Part
Greenberg, Robert
Role
Role Term: Text
reader
Name: Personal
Name Part
Nillni, Eduardo
Role
Role Term: Text
reader
Name: Corporate
Name Part
Brown University. Division of Biology and Medicine. Pathobiology
Role
Role Term: Text
sponsor
Genre (aat)
theses
Abstract
Trypanosomes are the causative agent of human African sleeping sickness and nagana in cattle. The bovine parasite, Trypanosoma brucei brucei (T. b. brucei), is unable to infect humans because of a small subclass of High Density Lipoprotein (HDL) known as trypanosome lytic factor (TLF). TLF is composed of several proteins including apolipoprotein A-1 (ApoA-1), apolipoprotein L-1 (ApoL-1), and haptoglobin-related protein (Hpr). Both ApoL-1 and Hpr have been shown to be trypanolytic. When these two proteins are reconstituted into an HDL particle, the lytic activity increases 100-fold, indicating that the proteins have a synergistic relationship. Furthermore, Hpr and ApoL-1 co-purify from plasma when an antibody to either Hpr or ApoL-1 is used, suggesting that the proteins co-assemble. Our findings suggest, however, that Hpr and ApoL-1 do not have direct interactions. To understand how Hpr and ApoL-1 are co-assembled, we have examined the biogenesis of TLF in human liver cells. Hpr and ApoL-1 localize to the surface of liver cells. Addition of HDL to the media results in scavenging of Hpr. We propose that Hpr and ApoL-I traffic to the cell surface of liver cells, where they are scavenged by HDL. Once co-assembled, Hpr and ApoL-1 containing HDL are still not lytic in vitro. Only after TLF binds Hb is TLF "armed" for killing. We show that Hb is required for binding and endocytosis of TLF. In addition, an iron chelator and free radical scavenger inhibit lysis by TLF-Hb. Using 500 kDa dextrans, we also show that TLF-Hb causes lysosomal membrane breakdown. Taken together, TLF-Hb generates free radicals by an iron dependent mechanism. The free radicals lead to lipid peroxidation and membrane breakdown. We now have a model of TLF that begins with its biogenesis and ends with its killing of trypanosomes. Hpr and ApoL-1 are assembled into HDL by a cell-mediated process. Upon infection by T. b. brucei, an acute phase response causes severe hemolysis. Hpr binds hemoglobin, activating the toxin. TLF is then endocytosed by the parasite, leading to cell death and clearance of the infection.
Subject (Local)
Topic
Trypanosome
Subject (Local)
Topic
Trypanosome lytic Factor
Subject (FAST) (authorityURI="http://id.worldcat.org/fast", valueURI="http://id.worldcat.org/fast/955053")
Topic
Hemoglobin
Record Information
Record Content Source (marcorg)
RPB
Record Creation Date (encoding="iso8601")
20091218
Language
Language Term: Code (ISO639-2B)
eng
Language Term: Text
English
Identifier: DOI
10.7301/Z0WS8RGG
Access Condition: rights statement (href="http://rightsstatements.org/vocab/InC/1.0/")
In Copyright
Access Condition: restriction on access
Collection is open for research.
Type of Resource (primo)
dissertations