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Proteomic and Functional Characterisation of LILRA3: Role In Proteomic and Functional Characterisation of LILRA3: Role in Inflammation Terry Hung-Yi Lee A thesis submitted in fulfilment of the requirements for the degree of Doctor of Philosophy Faculty of Medicine The University of New South Wales 2014 PLEASE TYPE THE UNIVERSITY OF NEW SOUTH WALES Thosis/Dissertatlon Sh09t Surname or Fam1ly name L l .... f.- First name / 'f (Z (1. 1 Other name/s H "'" ~ - \'I Abbre111at1on lor degree as given in the University calendar. Pt> .fh •I "!.1 '.j 17 J/ 0 School f t. t,...., ..> I <>f lVI e.~, u1 f <J t.r-UJ Faculty: M e. o1; c.• " e Tlrte Pr,te.;) ""l(. (j. f' il ,: ..... d .;" "' u"'"r &- d·er;..c&.L J .<\ ~ l/<.tf. Al: ~ " ' ~ ..... , ...., ~~~"""'" " "'~" Abstract 350 word s ma•imum: (PLEASE TYPE) Se e (f'JicJE' -------- Declaration relating to disposition of project thos tsldissortation I hereby grant to the Un1vers1ty of New South Wales or rts agents the nght to areh1ve and to make avatlabte my theSIS or dissertation '" whole or 1n part 1n the University libranes in all forms of media. now or here after known. subject to the provisions of the Copyright Act 1968 I retain all property nghts such as patent nghts I also retain the right to use in future works (such as artldes or bOOks) all or pan of this thesis or dlssenat1on I also authonse Un1vers1ty Microfilms to use the 350 word abstract of my thesis In Dissertation Abstracts International (this is appticable to doctoral ~~· ~l.A I Po .~.. <j 'I fJs JV VJignatur~ J1 Witness Date The Un1vers1ty recogn•ses that there may be exceptional circumstances requinng restrictions on copying or conditions on use. Requests for restnct1on for a penod of up to 2 yea rs must be made in wntlng. Requests for a longer penod of restriction may be considered in exceptional Circumstances and reQuire the aporoval of the Dean of Graduate Research. FOR OFFICE USE ONLY Date of completion of requirements for Award· THIS SHEET IS TO BE GLUED TO THE INSIDE FRONT COVER OF THE THESIS COPYRIGHT STATEMENT 'I hereby grant the University of New South Wales or its agents the right to archive and to make available my thesis or dissertation In whole or part In the University libraries in ail forms of media, now or here after known, subject to the provisions of the Copyright Act 1968. I retain ail proprietary rights. such as patent rights. I also retain the right to use in Mure works (such as articles or books) all or part of this thesis or dissertation. 1 also authorise University Microfilms to use the 350 word abstract of my thesis in Dissertation Abstract International (this is applicable to doctoral theses only). I have either used no substantial portions of copyright material in my thesis or I have obtained permission to use copyright material: where permiSSIOn has not been granted 1have applied/INiil apply for a partial restriction of the digital copy of my thesis or dissertation: Signed ...... b ..~ .... ~ .................. .. Date _l /4 {IS_ ............ .... .. AUTHENTICITY STATEMENT 'I certify that the Library depoM digital copy is a direct tl(luivatent of the final officially approved version of my thesis. No emendation of content has occurred and if there are any minor variations in formatting. they are the resun of the ::""''~~~ ~ 1M Date .. .... .. ..... .... ~. ..l.~.{. ..l .f. ... .. .. ... .............. .. ORIGINALITY STATEMENT 'I hereby declare that this submission is my own work and to the best of my knowledge it contains no materials previously published or written by another person, or substantial proportions of matenal which have been accepted for the 3ward of 3ny other degree or diploma at UNSW or any other educational institution, except where due acknowledgement is made in the thesis. Any contribution made to the research by others. with whom I have worked at UNSW or elsewhere, is explicitly acknowledged in the thesis. I also declare that the intellectual content of this thesis is the product of my own work, except to the extent that assistance from others in the project's design and conception or :,:: p~~'':;tk)'~:fit'':-"' Date ..........~ ... 1. ..~/ . !. J... ................ ............. Table of Contents TABLE OF CONTENTS .................................................................................................. I LIST OF TABLES ..................................................................................................... VIII LIST OF FIGURES ....................................................................................................... X ABBREVIATIONS .................................................................................................. XXV PUBLISHED WORK FROM THIS THESIS ................................................................. XXIX ACKNOWLEDGMENT ............................................................................................ XXX ABSTRACT .......................................................................................................... XXXI 1 INTRODUCTION ............................................................................................. 1 1.1 Overview ........................................................................................................... 1 1.2 Leukocyte immunoglobulin-like receptors (LILRs) ............................................ 3 1.2.1 Nomenclature, structure and classification .................................................. 3 1.2.1.1 Activating LILR (LILRA) ............................................................................... 7 1.2.1.2 Inhibitory LILR (LILRB) ............................................................................... 8 1.2.1.3 Soluble LILRs .............................................................................................. 9 1.2.2 Expression and functions of LILRs ............................................................... 11 1.2.2.1 Messenger RNA and/or protein expression ............................................ 11 1.2.2.2 Functions of LILRs .................................................................................... 11 1.3 Paired immunoglobulin-like receptors (PIRs) ................................................. 15 1.3.1 Groups of PIRs ............................................................................................. 15 1.3.1.1 Activating PIR (PIRA) ............................................................................... 17 1.3.1.2 Inhibitory PIR (PIRB) ................................................................................ 18 1.3.2 Expression and functions of inhibiting PIR (PIRB) ....................................... 19 1.3.2.1 Expression of PIRB ................................................................................... 19 i 1.3.2.2 Functions of PIRB .................................................................................... 20 1.4 Human LILRs and the murine orthologue PIRs ............................................... 21 1.4.1 Ligands for LILRs and PIRs ........................................................................... 21 1.4.1.1 MHC-I molecules as ligands .................................................................... 21 1.4.1.2 Non-MHC-I molecules as ligands ............................................................ 26 1.4.2 Downstream mechanisms of LILRs and PIRs ............................................... 30 1.4.2.1 Mechanisms of ITAM-mediated activating signalling ............................. 30 1.4.2.2 Mechanisms of ITIM-mediated inhibitory signalling .............................. 32 1.5 Nogo (reticulon 4 (RTN4)) ............................................................................... 36 1.5.1 Nogo and it isoforms ................................................................................... 36 1.5.2 Nogo functional domains ............................................................................ 38 1.5.3 Expressions and functions of Nogo ............................................................. 40 1.5.3.1 Expression of Nogo-A .............................................................................. 40 1.5.3.2 Functions of Nogo-A ................................................................................ 41 1.5.3.3 Expression of Nogo-B .............................................................................. 43 1.5.3.4 Functions of Nogo-B ................................................................................ 43 1.5.4 Receptors for Nogo ..................................................................................... 48 1.5.4.1 Nogo receptor (NgR) ............................................................................... 48 1.5.4.2 Nogo-B specific receptor (NgBR) ............................................................. 51 1.6 Glycosylation as important post-translational modification .......................... 52 1.6.1 Classes of protein glycosylation .................................................................. 53 1.6.1.1 N-glycosylation ........................................................................................ 53 1.6.2 The role of protein glycosylation in protein folding and ligand receptor interactions ................................................................................................. 55 1.6.3 Defective glycosylation and disease association ........................................ 57 1.7 Hypothesis and Aim ........................................................................................ 58 2 PRODUCTION AND CHARACTERISATION OF RECOMBINANT LILRA3 .............. 61 2.1 Introduction ...................................................................................................
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