I= AMINO ACID SEQUENCE OF THE HEMOGLOBIN FROM THE REPTILE UROMASTIX HARDWICK II
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Title of Thesis
AMINO ACID SEQUENCE OF THE HEMOGLOBIN FROM THE REPTILE UROMASTIX HARDWICK II

Author(s)
Sabira Naqvi
Institute/University/Department Details
H.E.J Research Institute Of Chemistry/ University Of Karachi
Session
1984
Subject
Chemistry
Number of Pages
81
Keywords (Extracted from title, table of contents and abstract of thesis)
amino acid, hemoglobin, uromastix hardwick ii, peptides, respiratory function, reptilian hemoglobin, cm-cellulose

Abstract
This is the first study on the hemoglobin of Uromastix Hardwick. We are reporting the amino acid sequence of its hemoglobin.

The separation of hemoglobin components and polypeptide chains were carried out on various columns and purity could not be achieved. The globin was, therefore carboxmethylated and cleaved with trypsin. The tryptic peptides were separated by gel filtration and further purified by reverse phase-high performance liquid chromatography. The amino acid composition of the peptides were determined after hydrolysis. The amino acid sequence of purified peptides was determined manually by the Adman degradation and DABITC method. The sequence of the peptides in the α and β chain was deduced from their homology with known sequence of ht polypeptides chains of other species. The sequence of Uromastix α- chain is compared with the α- chain of crocodile, viper, human and chicken, while the β-chain is compared with crocodile , human and chicken β-chain. The results show that hemoglobin chains are homologous with those of birds and mammals and the relationship hardly appear closer to known sequences of reptiles.

The results also show chain heterogeneity because at least two types of both α and β chain are present in the hemoglobin studies

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S. No. Chapter Title of the Chapters Page Size (KB)
1 0 Contents
41.47 KB
  0.1 Acknowledgement
  0.2 Summary
2 1 Introduction 1
151.68 KB
  1.1 General Description 3
  1.2 Respiratory Function Of Hemoglobin 4
  1.3 Hemoglobin Structure 4
  1.4 Evolution And Phylogeny 7
  1.5 Reptilian Hemoglobin 10
  1.6 Objective Of Present Studies 15
3 2 Experimental 17
191.98 KB
  2.1 Isolation Of Hemoglobin 17
  2.2 Photometric Estimation Of Hemoglobin Concentration 17
  2.3 Removal Of Heme 18
  2.4 Separation On Gel Chromatography 18
  2.5 Separation Of Cm- Cellulose With Salt Gradient 18
  2.6 Polyacrylamide Disc Gel Electrophoresis 20
  2.7 Reverse Phase- Hplc Goblin Chain Separation 23
  2.8 Chromatography On Cm-Cellulose For Goblin Chain Separation 25
  2.9 Carboxymethylation 26
  2.10 Cleavage With Trypsin 26
  2.11 Gel Chromatography Of Tryptic Peptides 27
  2.12 Digestion With Cyanogens Bromide 29
  2.13 Gel Chromatography Of Cyanogens Bromide Peptides 29
  2.14 Amino Acid Analysis 30
  2.15 N-Terminal Sequence Analysis By Dansylation Method 31
  2.16 N-Terminal Sequence Analysis By Edman Degradation 32
  2.17 Identification Of Pth - Amino Acids 34
  2.18 N-Terminal Sequence Analysis By Dabitc – Method 35
4 3 Results 38
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  3.1 Separation On Sephadex G-75 38
  3.2 Characterization Of Hemoglobin With Electrophoresis 38
  3.3 Separation Of Hemoglobin Ion-Exchanger 42
  3.4 Fractionation Of Globin Preparations 42
  3.5 Separation And Purification Of Tryptic Peptides 44
  3.6 Separation And Purification Of Cnb Peptides 55
  3.7 Amino Acid Composition Of Tryptic 55
  3.8 Sequence Of Tryptic Peptides 61
  3.9 Sequence Of Cyanogen Bromide Peptides 63
5 4 Discussion 65
88.89 KB
6 5 References 75
64.79 KB