I= AMINO ACID SEQUENCE OF HEMOGLOBIN COMPONENT FROM COLUMBA LIVIA (GERY PIGEON)
Pakistan Research Repository Home
 

Title of Thesis
AMINO ACID SEQUENCE OF HEMOGLOBIN COMPONENT FROM COLUMBA LIVIA (GERY PIGEON)

Author(s)
Chand Sultana
Institute/University/Department Details
H.E.J Research Institute Of Chemistry/ University Of Karachi
Session
1998
Subject
Chemistry
Number of Pages
157
Keywords (Extracted from title, table of contents and abstract of thesis)
amino acid, hemoglobin, columba livia, gery pigeon, columbiformes, globin, peptides, mammals, birds, reptiles, avian hemoglobin, respiration, heme

Abstract
The primary structure of hemoglobin from Grey Wild Pigeon (Columba livia) has been investigated. This is the first sequence determination study of the order Columbiformes.

Native globin was separated on ion exchanger and reversed phase high performance liquid chromatography. The intact subunits were further degraded by enzymatic and chemical cleavage into peptides. The structure of hemoglobin was established by sequence determination of tryptic peptides manually as well as automatically on Gas-phase sequencer. Homologous alignment of the structure with twenty other bird species belonging to different order indicate many interesting points. Among the different exchanges, positions α24 Leu, α26 Gly , α32 Leu, α64 Glu , α111 Ala, α113 Phe and α129 Val, β14, Ile, β55 ser, β59Asn, β61 Lys and β83 Gly are unique to pigeon hemoglobin. The various exchanges are discussed with reference to physiology closer to the order Anseriformes. On the basis of sequence data the phylogenic placement of Columba livia could be established among the birds having single hemoglobin component. The evolutionary relationship between mammals, birds and reptiles (Crocodiles) is discussed.

Download Full Thesis
2037.73 KB
S. No. Chapter Title of the Chapters Page Size (KB)
1 0 Contents
132.46 KB
2 1 Introduction 1
735.16 KB
  1.1 General Description 1
  1.2 Respiratory Proteins 1
  1.3 Avian Hemoglobin 10
  1.4 Heterogeneity In Avian Hemoglobin 30
  1.5 Oxygen Affinity Of Avian Hemoglobin 31
  1.6 High Altitude Respiration In Birds 35
  1.7 Expression Of Α d - Chain In Avian Hemoglobin 36
  1.8 Evolution And Phylogeny 38
  1.9 Objective Of The Work 40
3 2 Experimental 41
70.45 KB
  2.1 Blood From Grey Wild Pigeon 41
  2.2 Isolation Of Hemoglobin 41
  2.3 Photometric Estimation Of Hemoglobin 41
  2.4 Separation On Heme 42
  2.5 Separation By Gel-Chromatography 42
  2.6 Carboxymethylation Of Globin 42
  2.7 Separation By Ion Exchange Chromatography 42
  2.8 Separation Of Globin Chains By Reversed Phase High Performance Liquid Chromatography 43
  2.9 Polyacrylamide Gel Electrophoresis 44
  2.10 Cleavages 44
  2.11 Separation Of Tryptic Peptides On Reversed Phase High Performance Liquid Chromatography 45
  2.12 Separation Of Asp-Pro Peptides 45
  2.13 Purification Of Α- Peptides 46
  2.14 Amino Acid Analysis 46
  2.15 N-Terminal Sequence Analysis By Dansylation Method 46
  2.16 N-Terminal Sequence Analysis By Dabtic’s Method 47
  2.17 Automatic Sequence Analysis On Gas-Phase Sequencer 47
4 3 Results 48
227.78 KB
  3.1 Component Separation By Gel-Permeation Chromatography 48
  3.2 Characterization On Electrophoresis 48
  3.3 Separation Of Globin Chains By Ion-Exchange Chromatography 48
  3.4 Separation Of Globin Chains On Rp-Hplc 53
  3.5 Cleavage With Trypsin And Separation Of Tryptic Peptides 53
  3.6 Amino Acid Composition Of Chains And Peptides 53
  3.7 Chemical Cleavage Of Β-Chains 62
  3.8 Purification Of Α Peptides 62
  3.9 N-Terminal Determination By Dabitc ’s Method 62
  3.10 Automatic Sequence Determination Of Chains And Peptides 66
5 4 Discussion 68
903.99 KB
  4.1 Classification And Biology 68
  4.2 Primary Structure Of Α And Β-Chains 68
  4.3 Physiology 92
  4.4 Evolution 96
  4.5 Phylogenetic Placement Of Pigeon ( Columba Livia ) With Respect To Birds Human And Reptiles 115
  4.6 Conclusion 134
6 5 Appendix 139
55.97 KB
  5.1 Automatic Method Of Sequence Determination 139
  5.2 Edman Degradation 141
7 6 References 145
118.1 KB