Thaaw 6A fin that bovine spongiform phalopathy (BSE) has been transferred patients ro give chem new variant GieuefeldtJakob disease (CJD), which is ariably fatal. Ie presumed thac the of he infection was contaminated Polymorphism at codon 129 in the gman prion gene encoding either Met ot i has been identified and might bate © CJD suscepebilcy or ton time. The urman distribution ig. Caucasians is 37% Met/Met, 51% Nal and 12%Val/Val. All the cases of je variant CJD so far reported ate Piorygous for Met, so it remains posible those expressing Val at codon 129 ght occur in the fature and che icopathologic phenotype might be erent from the cases already tntire complement of proteins duced in a cll or organism is called proteome, derived from the genome, this the entire complement of genetic formation. To study the proteome, ih might well comprise hundreds or jan of different proteins it has been ry to develop large-scale and high- ghput procein analysis techniques, the techniques essentially define the € of proceomics. The most n echnique is 2D-gel horess. A protein mixture is Toads 02 polyacrylamide gel and the proteins gated by isoclectric focusing, which ens the proteins migrate in a pH. ent until they reach theit isoelectric in: The gel is chen equilibrated in a get (ee p. 12) and the proteins pasted in the second dimension toning to their M,, The gel is then ined and the proteins revealed asa een of spots The structure of the ins can be determined by such thods as mass spectrometry. The ces heeween proteomes can be de, the difference between the omes of liver and hepatoma, and ein disease markers can be devised. example ofthe use of proteomics is differentiation berween types of ler cancer, which can be difficult to guish by histological examination, Western world, bladder cancers are uly transitional cell carcinomas, whereas lose in Afica are often squamous cell cinoms. The techniques are also useful the pharmaceutical industry to check the toxic effects of drugs ‘STRUCTURE AND PROPERTIES OF MYOGLOBIN AND HEMOGLOBIN STRUCTURE OF MYOGLOBIN globin involves eight helical regions as shown in Fig, 4.11 These are denoted by the letters A-Hs the regions joining the helices are denoted by. pairs of leters (e.g. PG, joining helices F and G, not shown in the figure) ‘The architecture of my Histdines in helices E and F inceraet with heme on either side. The oxygen (O:) molecule sits at W. Helices E and F form the walls of a box for the heme; B, G and H are the floor and the CD corner closes the open end. The heme pocket consists in the main of non-polar (group 1) amino acids Fig. 4.11 The architecture of myoglobin involves eight helical regions. The histidines interacting with heme are shown in ree. Fig. 4.12 The artangement of the four chains of hemaglobin, Myoglobin carries charged arnino acid residues at positions that are hydrophobic in hemoglobin and which are imporcane in bonding the hemoglobin subunits cogether, 28 will be explained, For this reason myoglobin molecules do not associate together STRUCTURE OF HEMOGLOBIN ‘Comparison of subunits with myoglobin Hemoglobin (Fig. 4.12) consists of four chains comprising two pairs of identical chains; these are designated 01; and BB» ‘There are few bonds between the evo chains or berween the two chains However, there are strong hydrophobic 6bonds berween unlike chains (eg 0B» 048s, 0B oF OsB).The bonds between an Gand a B monomer are stronger than ca or B-B bonds or those between one OB dimer and another. Thu, if hemoglobin is dissociated by tweatmene with 8-M urea, {9B dimers always result rather than at oF BB dimers, Tis shown in Fig. 4.13. ‘The a: chain of hemoglobin differs from the B chain by the deletion of one residue inthe NA segment, the addition of «wo residues in che AB corner and the deletion of six tesidves in the CD segment and the D helix “The binding of oxygen to hemoglobin results in the movements of and chains a anit relative to the ob and Bs chains. Deoxyhemoglobin is denoted as having the"T (tense) form, and oxyhemoglobin the R. (relaxed) form. The formation of the quaternary structure of hemoglobin “This is shown in Fig. 4.145 the four chains interact to give a compact structure, which is essential for correct protein function. If there is any change in the primary structure of the chains that affects the {quaternary structure the properties of the hemoglobin will be changed. Although this folding is common to the hemoglobin of ‘every species possessing this type of mily about ten amino acid residues are invariant (Le. present in every case). Some changes in the prim: seructure make lice difference to the tertiary structure, whereas others have a molecule, profound effect, as in sickle-cell hemoglobin (Gee below) HEMOGLOBIN POLYMORPHISM IN THE HUMAN In the normal human Figure 4.15 shows the different types of hemoglobin that are present in the normal human and which arise ffom the expresion of the gene loci. In particular, it should be noted that fetal hemoglobin differs fom adult hemoglobin. CLINICAL IMPLICATIONS ~ HEMOGLOBINOPATHIES “The inherited disorders of hemoglobin fall into three groups. First, there are the seructural hemoglobin variants deseribed below Second, there are the thalasemias which are characterized by a reduced ra of production of either the & or B globin chains and which ate, therefore, divided into the er and B thalassemias, Finally, chere 62 Chains 1G, tere Hemogiobing Sete Hp Poriane Ho Gower 1 Hb Gower 2 Embryo oe Hoa, Hoa Fig 4.13 The dso fof hemoglobin by urea ‘causes the formation ‘mixed: (uit often associtod with I plasms ie used instoad of serum, fibrinagen bend gives the appearance ‘ofa parapratln leaing to ‘A monoclonal band, refered to 363 paraproin which du to produc: fiom ofa specie immunogiobuin by 8 malignant clone of calls Pareprotins fd sometimes globulin ‘A polyclonal gammapathy with inmany ferent Inmmonoglabulins of al castes Elevated «and, petsins. Usually Se iui, So-caled Fig. 5.1 Electrope ‘of serum prateins on cellulose acetate membrane using @ buffer at pH 8.6 can Used to diagnose abnormalities in a patient's scum, The protein bands ae Primary Impaired synthesis of Inman Immunoglobulins ‘steieney Unvaty arta Mute : Ur sao found roel ia ether esoar3s Nephvosc Albumin ost into win yndrome soda chemosis le Infection decreased a chronic | lymphatic eased y-globulin footw seeteet ram tae I | j | Aasieypsin fsterencr “able 5.1 The principal constituents of the serum globulin bands separated by ‘electrophoresis Globulin Representative constituents a ‘Thyroxine-binding globulin Transcortin Glycoprotein Lipoprotein anttrypsin % Haptoglobin Giycoprotein Macroglobulin {ceruloplasmin p ‘Transferrin Lipoprotein Glycoprotein 1 2620, 96 9A tmisieacing diagnos’ ‘acanteypsn dficieney ‘Saocitea win emphysema ofthe ting in adults, ena juvenile Visualized by staining witha dye, Table 5.4 lists the principal protein constituents the various bands of ‘lobulin. ALS, albu These are listed in Table 5.2. More information about the significance of so of the proteins will be found later inthe text. Diagnosis is also assisted by the measurement of various enzymes in the settm, as explained in Table 6.1 (p. 89) ‘Thyroxine-binding globulin (TBG) cat 70% of the thyroid hormone (T, and.) in plasma. About 2 alburnin and the testis bound t0 transthyretin, formerly known as pre- ‘may transform into amyloid fibrils that dleposit in cardiac and other tissues, giving busin. Four transthyretin molecules rise to the condition known as senile systemic amyloidosis in some elderly individuals, Patients suffering from fami amyloidotic polyneuropathy inherit a single-point mutation, valine to ‘methionine, which causes the soluble, lobular eransthyretin to polymerize and ‘gradually deposit a fibrous material in the