NUCLEIC ACIDS. An INTRODUCTION. Two classes of Nucleic Acids
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1 NUCLEIC ACIDS An INTRODUCTION Two classes of Nucleic Acids Deoxynucleic Acids (DNA) Hereditary molecule of all cellular life Stores genetic information (encodes) Transmits genetic information Information on protein sequence is encoded in DNA and translated by the help of messenger Ribonucleic Acid (mrna) Ribonucleic Acids Encode and Translate information on DNA to proteins 1
2 Composition and Structure of Nucleic Acids Nucleobases Derivatives of purine and pyrimidine bases Sugars Phosphoric acid DNA nucleobases Adenine (A) and Guanine (G) Tymine (T) and Cytosine (C) RNA nucleobases A and G Uracyl (U) and Cytosine (C) Nucleobases Uracyl thymine without the methyl group DNA nucleobases Adenine (A) and Guanine (G) Tymine (T) and Cytosine (C) RNA nucleobases A and G Uracyl (U) and Cytosine 2
3 Sugars β-d-ribose and β-d-deoxyribose DNA Nucleosides Base (N9 or N1 atom) + sugar (C1 atom) N-glycosidic bond 3
4 Building Blocks: Nucleotides Nucleoside + phosphate group(s) 5 -hydroxylic group of sugar phosphate Ester bond Polynucleotides A T G C sugar phosphate sugar phosphate sugar phosphate sugar... Backbone : sugar + phosphate Side groups: nucleobases Reading from From Nucleotide with free phosphate group (5 end) To nucleotide with free 3 hydroxyl group (the 3 end) 4
5 Base Pairing Double Strand Formation Double strand DNA is inherently very sensitive/ fragile Strands Pairing 5
6 The Double Helix Denatured during replication and mrna synthesis Denatured during PCR (amplification of DNA) Levels of Structural Organization Primary Sequence of nucleotide Name of nucleobases used : ATCT Secondary Double stranded helix structure Tertiary Folding of the double helix Example: super coiled 6
7 Structure of RNA Single strand Folds on itself thus allowing pairing of bases Synthesis of Proteins Procaryote cell Eucaryote cell 7
8 Synthesis of Proteins Synthesis of Proteins 1. Transcription One strand of the DNA double helix is used as a template by the RNA polymerase to synthesize a messenger RNA (mrna). mrna migrates from the nucleus to the cytoplasm. mrna goes through different types of maturation including one called splicing when the non-coding sequences are eliminated. The coding mrna sequence can be described as a unit of three nucleotides called a codon. 2. Translation The ribosome binds to the mrna at the start codon (AUG) that is recognized only by the initiator trna. The ribosome proceeds to the elongation phase of protein synthesis. During this stage, complexes, composed of an amino acid linked to trna, sequentially bind to the appropriate codon in mrna by forming complementary base pairs with the trna anticodon. The ribosome moves from codon to codon along the mrna. Amino acids are added one by one, translated into polypeptidic sequences dictated by DNA and represented by mrna. At the end, a release factor binds to the stop codon, terminating translation and releasing the complete polypeptide from the ribosome. One specific amino acid can correspond to more than one codon. The genetic code is said to be degenerate. 8
9 Synthesis of Proteins: Genetic Code Protein Synthesis Folding, disulfide bridges formation, posttranslational modification Post-translational modifications Not determined by DNA sequence Cleavage of amino acids Phosphorylation Acetylation glycosylation 9
10 Analytical Challenges for the Analysis of Biological Macromolecules Inherent sensitivity Both proteins and nucleic acid 3D structure are very sensitive to environmental conditions such as ph, ionic strength, solvent composition Why? stabilized by H-bonds, other dipole-dipole interactions, electrostatic interactions, van der Waals interactions Problems Denaturation Crystallization challenges Analytical Challenges for the Analysis of Biological Macromolecules Size of macromolecules Several signals that must be resolved and analyzed Complex three dimensional Structure NMR challenges Number of macromolecules in real samples Separation methods Methods for selective detection Amount of sample Sensitivity must be high Amplification 10
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