maenas (intertidal zone) and Segonzacia mesatlantica - Station ...

More documents

Recommendations

Info

The Structural Analysis of Large Noncovalent Oxygen Binding Proteins Current Protein and Peptide Science, 2008, Vol. 9, No. 2 167 Table 3. Experimental Subunit Masses, Native Masses, Calculated Masses of Crustacean Hcys Species Ag. state Subunit Composition Native Mass Calculated Mass Expl Mass (Da) ±s.d. a Method [ref] Ag. state Expl mass (Da) ±s.d. Relative Abundance Method [ref] Cltd mass from mean subunit mass Difference % Pleocyemata Caridea Bresiliidae Rimicaris 6 74 816,2 ±1 ESI-MS 12 935 000 ±40 000 0,03 MALLS 905 060,8 3,20 exoculata 6 74 865,3 ±2,3 [165] 6 469 000 ±9 000 0,89 [105] 450 283,9 3,99 6 74 902,5 ±1,5 1 80 000 ±10 000 0,08 6 74 939,6 ±2,6 6 74 979,8 ±4,5 6/12 75 018,5 ±6 12 75 326,4 ±5,1 12 75 539,3 ±6,2 Brachyura Bythograeidae 6/12 75 008,3 ±1,2 ESI-MS 12 nd 0,45 905 212,65 Cyanagraea 6/12 75 062,2 ±4,2 [107] 6 0,55 [107] 452 173,4 praedator 6/12 75 110,3 ±2,2 6/12 75 321,6 ±1,3 6/12 75 375,9 ±6 12 75 419,1 ±5,3 6/12 75 533,1 ±1,1 12 75 628,6 ±3,8 Bythograea nd 74 454 ESI-MS 2 235 051 ±303 ESI-MS 2 242 410 0,33 thermydron 74 549 [62] 30 b 2 256 550 ±396 [62] 2 242 410 0,63 74 647 24 1 804 368 ±284 1 793 928 0,58 74 722 18 1 354 950 ±480 0,01 ESI-MSc 1 345 446 0,70 74 728 12 902 570 ±110 0,13 SEC (proportions) 896 964 0,62 6 450 310 ±260 0,67 [73] 448 482 0,41 1 74 999 ±85 0,18 Segonzacia nd 75 234 ESI-MS 18 1 364 968 ±264 ESI-MS 1 361 574 0,25 mesatlantica 75 322 [62] 1 347 998 ±300 [62] 1361574 1,01 75 541 12 909 628 ±157 907 716 0,21 75 967 6 453 889 ±45 453 858 0,01 Portunidae Carcinus nd 73 931 ESI-MS 18 1 374 136 ±280 ESI-MS 1 349 334 1,80 maenas 74 049 [62] 12 975 332 ±151 [62] 899 556 7,77 74 117 888 652 ±299 899 556 1,23 74 966 12 b 902 675 ±392 899 556 0,35 75 088 6 487 337 ±35 449 778 7,71 75 161 450 541 ±44 449 778 0,17 75 234 75 459 12 915 913 ±3 032 MALLS 899 556 1,79 75 519 6 466 631 ±10 110 personal observations 449 778 3,61 Dendrobranchiata Penaeoidea Penaeidae Penaeus nd 24d 1 800 000 MALLS nd monodon 12 950 000 [166] 6 nd a When subunit is present in dodecamer and hexamer, mean of dodecamer and hexamer determinations is given; given sd is the maximum sd of the determinations b Precise charge state assignment is ambiguous and two mean masses are possible c For mass data; similar results for 18,12,6 and 1-mer were obtained by [62] d The 24-mer is a minor component; the major component is the hexamer 88
168 Current Protein and Peptide Science, 2008, Vol. 9, No. 2 Bruneaux et al. weight; retention times do not vary between different hemolymph samples (b) Masses obtained for LtHb. Values for MALLS is taken from [32]. Partially native value for ESI-MS is based on the native mass obtained by [63] for the dodecamer; it is calculated for 12 dodecamers and 36 linkers. The linker average mass is calculated from [23] data, with abundances from [167] (27 659.94 Da). Values for other methods are taken from [116] and include LS, STEM, EM and SE determination. The model mass was taken from [32]. (c) Masses obtained for Arenicola marina HBL-Hb. MALLS values are from [89]. Partially native ESI-MS values were calculated from native dodecamer mass from [64] and from the average linker dimer mass calculated from [83] (50 945 Da). It is calculated for 12 dodecamers and 18 linker dimers. Values for other methods are taken from [32], considering only data posterior to 1950. They includes SE, SD and SAXS. The model mass was calculated from the dodecamer mass calculated by [71] from denaturing ESI-MS data and from the average linker dimer mass calculated from [83]. It should be noted that the partially native ESI-MS determination for HBL-Hb masses shares the linker data with the calculated model mass. Fig. (4). Comparison between masses estimated by MALLS, ESI-MS and other methods for Hc and HBL-Hb. Bars are mean values with s.d.; the dashed line corresponds to a model calculated from masses obtained in denaturing ESI-MS (a) Masses obtained for Carcinus maenas Hc. The same samples (n=16) were analyzed by MALLS and ESI-MS except for one which could not analyzed by ESI-MS. The model mass was calculated for a dodecamer using an intensity-weighted mean subunit mass with two copper atoms of 74 826.91 Da, obtained from all samples in denaturing conditions. The SEC mass was obtained after calibration of a Superose 6 column by markers of known molecular HBL-Hb has been reported to involve independent folding of individual domains followed by domain interaction for the oligochaete Lumbricus terrestris Hb [110]. The stability of the quaternary structure of annelid HBL-Hbs has been studied by changing the chemical composition of the medium: by varying pH, incubation in the presence of chaotropic salts, or denaturing agents. There is an increasing interest in understanding the dissociation and association process of this Hb because it provides useful information about subunit interactions necessary to maintain the quaternary structure. Moreover, AmHb has been proposed as useful model system for developing therapeutic extracellular blood substitute [7, 8] and required a full study of subunit interactions in order to find the correct storage and transfusion buffer composition. In order to analyze the interactions between globin and linker subunits constituting HBL-Hb, dissociation and reassociation experiments were carried out under several conditions: exposure to urea and alkaline pH and the effect was followed by SEC-MALLS and ESI-MS. Thanks to the complementarity of information obtained with ESI-MS and MALLS analysis, Rousselot and collaborators have bring to light a novel and complex mechanism of dissociation for AmHb compared to other annelid extracellular Hbs studied to date [6]. Even though the chemically-induced dissociation triggers partial degradation of some subunits, spontaneous reassociation was observed to some extent. Parallel dissociation of LtHb under similar conditions shows striking differences that allow us to propose a hypothesis on the nature of the intersubunit contacts that are essential to form and to hold such a complex quaternary structure. 6.2. An Application Example: The Case of Arenicola marina HBL-Hb MALLS analysis of partially dissociated AmHb at slightly alkaline pH yielded profiles shown in Fig. (5), with molecular masses Mw (Fig. 5a) and gyration radius Rg (Fig. 5b) estimated during the elution profile at 3 different incubation times. The estimated Mw (Fig. 5a) decreases during incubation time and the polydispersity estimated by the molar mass slopes assumes a downward curvature shape, 89
Page 1 and 2:
THESE DE DOCTORAT DE L’UNIVERSITE
Page 3:
i Résumé / Abstract Réponse adap
Page 6 and 7:
iv SOMMAIRE 4.2 Objectifs de l’é
Page 8 and 9:
2 CHAPITRE 1. INTRODUCTION Les éco
Page 10 and 11:
4 CHAPITRE 1. INTRODUCTION 1.1 Mili
Page 12 and 13:
6 CHAPITRE 1. INTRODUCTION Les prin
Page 14 and 15:
8 CHAPITRE 1. INTRODUCTION TAB. 1.1
Page 16 and 17:
10 CHAPITRE 1. INTRODUCTION et cycl
Page 18 and 19:
FIG. 1.5 - Localisation des princip
Page 20 and 21:
(a) FIG. 1.6 - Fonctionnement d’u
Page 22 and 23:
16 CHAPITRE 1. INTRODUCTION TAB. 1.
Page 24 and 25:
18 CHAPITRE 1. INTRODUCTION FIG. 1.
Page 26 and 27:
20 CHAPITRE 1. INTRODUCTION Différ
Page 28 and 29:
22 CHAPITRE 1. INTRODUCTION FIG. 1.
Page 30 and 31:
24 CHAPITRE 1. INTRODUCTION le mili
Page 32 and 33:
26 CHAPITRE 1. INTRODUCTION Capacit
Page 34 and 35:
28 CHAPITRE 1. INTRODUCTION TAB. 1.
Page 36 and 37:
30 CHAPITRE 1. INTRODUCTION 700 600
Page 38 and 39:
32 CHAPITRE 1. INTRODUCTION férent
Page 40 and 41:
34 CHAPITRE 1. INTRODUCTION (a) (b)
Page 42 and 43:
36 CHAPITRE 1. INTRODUCTION (a) (b)
Page 44 and 45: 38 CHAPITRE 1. INTRODUCTION (a) (b)
Page 46 and 47: 40 CHAPITRE 1. INTRODUCTION 1 P 50
Page 48 and 49: 42 CHAPITRE 1. INTRODUCTION presque
Page 50 and 51: 44 CHAPITRE 1. INTRODUCTION affecte
Page 52 and 53: 46 CHAPITRE 1. INTRODUCTION FIG. 1.
Page 54 and 55: 48 CHAPITRE 1. INTRODUCTION FIG. 1.
Page 56 and 57: 50 CHAPITRE 1. INTRODUCTION fure co
Page 58 and 59: 52 CHAPITRE 1. INTRODUCTION amélio
Page 60 and 61: 54 CHAPITRE 1. INTRODUCTION 1.5 Que
Page 62 and 63: 56 CHAPITRE 2. ESI-MS ET MALLS APPL
Page 78 and 79: The Structural Analysis of Large No
Page 108 and 109: 102 CHAPITRE 2. ESI-MS ET MALLS APP
Page 110 and 111: 104 CHAPITRE 3. STRUCTURE DE L’HC
Page 140 and 141: 134 CHAPITRE 4. PLASTICITÉ PHÉNOT
Page 142 and 143: 136 CHAPITRE 4. PLASTICITÉ PHÉNOT
Page 144 and 145:
138 CHAPITRE 4. PLASTICITÉ PHÉNOT
Page 146 and 147:
Page 148 and 149:
Page 150 and 151:
Page 152 and 153:
Page 154 and 155:
Page 156 and 157:
Page 158 and 159:
Page 160 and 161:
Page 162 and 163:
Page 164 and 165:
Page 166 and 167:
Page 168 and 169:
Page 170 and 171:
Page 172 and 173:
Dénaturant Natif
Page 174 and 175:
Page 176 and 177:
Page 178 and 179:
172 CHAPITRE 5. ADAPTATIONS RESPIRA
Page 180 and 181:
Page 182 and 183:
Page 184 and 185:
Page 186 and 187:
Page 188 and 189:
Page 190 and 191:
Page 192 and 193:
Page 194 and 195:
Page 196 and 197:
Page 198 and 199:
Page 200 and 201:
Page 202 and 203:
Page 204 and 205:
Page 206 and 207:
Page 208 and 209:
Page 210 and 211:
Page 212 and 213:
Page 214 and 215:
Page 216 and 217:
Page 218 and 219:
Page 220 and 221:
Page 222 and 223:
216 CHAPITRE 6. CONCLUSION ET PERSP
Page 224 and 225:
Page 226 and 227:
Page 228 and 229:
Page 230 and 231:
Page 233 and 234:
Annexe A Détermination des masses
Page 235 and 236:
229 10000 h280 .M h) h280−h337 80
Page 237 and 238:
Annexe B Détermination des distrib
Page 239 and 240:
233 Modèle 2 : 2 sous-unités lég
Page 241 and 242:
235 1 0.9 0.8 6 s-u aléatoires 2 s
Page 243 and 244:
Bibliographie Adair, G. S. The hemo
Page 245 and 246:
BIBLIOGRAPHIE 239 Catalina, M. I.,
Page 247 and 248:
BIBLIOGRAPHIE 241 Farmer, C. S., J.
Page 249 and 250:
BIBLIOGRAPHIE 243 Hourdez, S. Adapt
Page 251 and 252:
BIBLIOGRAPHIE 245 Mangum, C. P. Sub
Page 253 and 254:
BIBLIOGRAPHIE 247 Rainer, J., C. P.
Page 255 and 256:
BIBLIOGRAPHIE 249 Svedberg, T. et I
Page 257 and 258:
BIBLIOGRAPHIE 251 Vanin, S., E. Neg
Page 259 and 260:
Liste des figures 1.1 Cycle des mar
Page 261:
LISTE DES FIGURES 255 4.17 Profils
Page 265 and 266:
Table des matières Résumé Sommai
Page 267 and 268:
TABLE DES MATIÈRES 261 4.3.5 Spect
Page 269 and 270:
TABLE DES MATIÈRES 263 Table des m
show all

maenas (intertidal zone) and Segonzacia mesatlantica - Station ...

Create successful ePaper yourself

Delete template?

Save as template?