IX
Zusammenfassung
Affinität zu Peptid-MHC-Klasse-II Komplexen mit geringer Affinität im Vergleich zu
Komplexen mit hoher Affinität. Diese Ergebnisse können genutzt werden, um Ko-
kristallisationsexperimente anzuleiten, die darauf gezielt sind, die Bindungsstelle des
kleinen Moleküls zu bestimmen. Die Information der spezifischen Interaktionen
zwischen J10 und MHC-Klasse-II Molekül könnte zur Verbesserung von J10 eingesetzt
werden, welches
Potential hat, therapeutisch eingesetzt zu werden.
X
List of publications
List of publications
Schulze, M.S., A.K. Anders, M.J. Call, K.W. Wucherpfennig: Structure of a human
MHC class II molecule with a partially filled peptide-binding groove. 2012,
(manuscript in preparation).
Schulze, M.S., K.W. Wucherpfennig: The mechanism of HLA-DM induced peptide
exchange in the MHC class II antigen presentation pathway.
Current Opinion in
Immunology. 2012, 24:105-111.
Anders, A.K., M.J. Call,
M.S. Schulze, K.D. Fowler, D.A. Schubert, N.P. Seth, E.J.
Sundberg, K.W. Wucherpfennig: HLA-DM captures partially empty HLA-DR
molecules for catalyzed removal of peptide.
Nature Immunology. 2011, 12:54-
61.
XI
List of abbreviations
List of abbreviations
ASU
-
asymmetric unit
CAPS
-
N-cyclohexyl-3-aminopropanesulfonic acid
CD
-
cluster of differentiation
CHO
-
chinese hamster ovarian
CIITA
-
class II transcriptional transactivator
CLIP
-
class-II-associated invariant
chain peptides
DMSO
-
dimethyl sulfoxide
DNP
-
dinitrophenyl
DTT
-
dithiothreitol
E. coli
-
Escherichia coli
ER
-
endoplasmatic reticulum
FP
-
fluorescence polarization
GM/CA-CAT
-
General Medicine and Cancer institutes Collaborative Access Team
HA
-
hemagglutinin
HLA
-
human leukocyte antigen
HPLC
-
high-performance liquid chromatography
HSQC
-
heteronuclear single quantum coherence
Ii
-
invariant chain
MBP
-
myeline basic protein
MES
-
2-morpholinoethanesulfonic acid
MHC
-
major histocompatibility complex
MHC I
-
major histocompatibility complex class I
MHC II
-
major histocompatibility complex class II
MLE
-
MHC loading enhancer
MOI
-
multiplicity of infection
MPD
-
2-methyl-2,4-pentandiol
MWCO
-
molecular weight
cut off
NE-CAT
-
Northeastern Collaborative Access Team
NMR
-
nuclear magnetic resonance
nOe
-
nuclear Overhauser effect
NOESY
-
Nuclear Overhauser effect spectroscopy
PAGE
-
polyacrylamide gel electrophoresis
PDB
-
protein data bank
PEG-x
-
polyethylene glycol with a mean molecular mass of x g/mol
RU
-
response unit
SDS
-
sodium dodecyl
sulfate
Sf
-
Spodoptera frugiperda
SPR
-
surface plasmon resonance
TFA
-
trifluoroacetic acid
tr
-
transverse relaxation-optimized
TROSY
-
transverse relaxation optimized spectroscopy
XII
Table of contents
Table of contents
Gutachter _______________________________________________________________ II
Declaration _____________________________________________________________ III
Acknowledgment ________________________________________________________ IV
Summary _______________________________________________________________ V
Zusammenfassung ______________________________________________________ VII
List of publications _______________________________________________________ X
List of abbreviations _____________________________________________________ XI
Table of contents ________________________________________________________ XII
1
Introduction .......................................................................................................... 1
1.1
The major histocompatibility complex ................................................................ 1
1.2
MHC class II dependent pathway of antigen presentation................................ 2
1.3
Structure and function of MHC class II/peptide complex and comparison
with MHC class I/peptide complex ...................................................................... 5
1.4
The role of HLA-DM in the MHC class II presentation pathway and
comparison to the peptide loading complex of MHC class I molecules ............ 9
1.5
Structure of HLA-DM and lateral interaction with HLA-DR ........................ 11
1.6
Proposed mechanisms for HLA-DM-catalyzed peptide exchange .................. 14
1.7
HLA-DO: a negative regulator of HLA-DM ..................................................... 16
1.8
MHC loading enhancers ..................................................................................... 17
1.9
Scopes and objectives of this study .................................................................... 21
2 Chapter I: Investigating enhanced peptide exchange of MHC class II molecules
by the small molecule catalyst J10 ........................................................................ 22
2.1
Introduction ......................................................................................................... 22
2.2
Materials and Methods ....................................................................................... 24
2.2.1
Preparation and crystallization of HLA-DR2/MBP in the presence of J10-1 and J10-12 .
....................................................................................................................................... 24
2.2.2
Preparation of isotope labeled MBP peptide ................................................................. 25
2.2.3
Preparation of HLA-DR2/peptide complexes ............................................................... 26
2.2.4
NMR experiments ......................................................................................................... 27
2.3
Results and discussion ......................................................................................... 28
