Wednesday, 16 December 2020

What is Protein, Structure of protein, Types of protein, Ramachandran plot

1. Introduction of protein

2. Structure of protein

3. Types of protein

4. Ramachandran plot of protein

5. Extraction and techniques 

 

Protein Introduction -

Proteins are  Nitrogenous macromolecules substances found in living system. Essential for cell structure and cell functions.

Protein term first suggested, in 1838, by a Swedish chemist Berzelius.

Macromolecules - Very high molecular weights, The term macromolecule was  coined by Schaudinger in 1920s.

Protein are polymers of The Amino acids.

 

In body -

                  H20 > Proteins > Lipids >  Carbohydrates > Nucleic Acid.

                             O2 - 60%,  C - 20%,  H - 10%,  N - 3%,  Ca - 2.5%.

Amino acids -

                       




R  =  H  ----  Glycine amino acid

R  =  CH3   ----  Alanine amino acid

R  =  CH2OH  ----  Serine amino acid.

 

 

22 Amino acids discovered in cells.

1. Arginine (R),

2. Histidine (H),

3. Isoleucine (I),

4. Leucine (L),

5. Lysine (K),

6. Methionine (M),

7. Phenylalanine (F),

8. Theonine (T),

9. Tryptophan (W),

10. Valine (V),

11. Alanine (A),

12. Asparagine (N),

13. Aspartic acid (D),

14.  Glycine (G),

15.  Serine (S),

16.  Tyrosine (Y),

17. Glutamic acid (E),

18.  Cysteine (C),

19. Glutamine (Q),

20. Proline (P),

21.  Selenocysteine (U),

22. Pyrrolysine (O)

 

         

  New discovered amino acids -

-   Selenocysteine (encode by UGA codon)

-   Pyrrolysine  (encode by UAG codon).

Amino acids nature -

          Acidic amino acids - Aspartic acid (D),    Glutamic acid (E).

          Basic amino acids - Lysine (K), Arginine (R), Histidin (H), Tryptophan (W).

          Neutral amino acids - Glycine (G), Valine (V), Alanine (A).

          Aromatic amino acids - Phenylalanine (F), Tyrosine (Y), Tryptophan (W).

 

 

Essential Amino Acids -

Essential Amino Acids cannot synthesized by de nevo, in organism at a rate commensurate with its demand.

Examples -

Arginine (R), Histidine (H), Isoleucine (I), Leucine (L), Lysine (K), Methionine (M), Phenylalanine (F), Theonine (T), Tryptophan (W), Valine (V).

 

Non essential amino acids -

Non essential Amino Acids synthesized by de nevo, in organism at a rate commensurate with its demand.

Examples -

Alanine (A), Asparagine (N), Aspartic acid (D), Glycine (G), Serine (S), Tyrosine (Y). Glutamic acid (E).

 

Conditional essential -

 Cysteine (C), Glutamine (Q), Proline (P).

 

Primary amine - One Carbon bounded with to the Nitrogen.

Second amine - Two Carbon bounded with to the Nitrogen.

Tertiary amine - Three Carbon bounded with to the Nitrogen.

 

Protein chain formation - Two amino acid attached with dipeptide bond.

                    


    Protein Molecular weight =  Add Amino acids molecular weights - 18 (H20, release).

 

Protein structure -

             



Effects of Temperature  on protein structure - high temperature least affects  primary structure.

Types of proteins -

Simple protein     -        1. Fibrous protein.    2. Globular protein.

Compound protein   -    1. Glycoprotein.   2. Leithoprotein.    3. Phoshpoprotein  etc.

 

Simple Protein -

Made by  only amino acids

1. Fibrous protein -

                                   Elongated, rod like structures, Form by strong hydrogen binding- between intermolecular strands, some time repeated unit, resistant to enzymatic digestion, Mostly- Primary & Secondary structures.

Examples - Collagen (Most abundant protein in animal body),

Elastin,

Keratin (Hardest protein, due to cysteine).

 

2. Globular protein -

                                   Spherical in shape, forms colloids with water, dissolve in water. Weak Hydrogen bonding, Enzymatic functions, mostly primary, secondary, tertiary and Quaternary structures.

Examples -

                   Rubisco (Most abundant protein on Earth / plants),

                   Albumin,

                   Globin,

                   Histone,

 

Notes - Fibrous proteins and globular proteins differ in size, shape, solubility, appearance as well as in function.

 

Compound protein -

                                     Made by Amino acids and non amino acids parts (Prosthetic part).

Examples -

Phoshpoprotein.    -         Amino acids + Phosphoric acids

Like - Milk protein (Casein),  Pepsin (digestive enzymes secret from stomach's  chief cells.

Leithoprotein  -    Amino acids + Lecithin

Like -  Fibrinogen (blood clotting protein)

Glycoprotein -      Amino acids + Carbohydrates

Like - LH (Leutinizing Hormone), FSH (Follicular stimulating factor,  Anterior pituitary Hormone).

Antibodies.

Fertilizin (egg surface protein),

Antuferilizing ( sperm surface protein).

 

Lipoprotein -    Amino acids + lipids

Like - plasma membranes

 

Nucleoprotein -   Amino acids + nucleic acids

Like -      DNA+ HISTONE PROTEIN,      

                       RNA + Protein

         Chromoprotein -  Amino acids + Metals

   Like -  Haemoglobin (Fe),

              Cytochrome (Fe).

 

Ramachandran plot -

Introduced by G. N. Ramachandran, Peptide conformations are defined by  the values of fi and sai.

For L amino acids (mostly found in eukaryotes, NH2 group presents in left side).   

Notes - Glycine ( Much broader range area)

              Proline (Greatly restricted area)

               For D amino acids ( NH2 group presents in right side), its are mirror form L amino acids.

 

"Amino acids encode by Codon (three nucleotides long unit)"

 

Isoelectric pH (pl)   -      Protein's Net electric charge is zero, but contain positive and negative charge groups. (Zwitterion).

                                  pH below its pI -    +Ve Change (Acidic)

                                   pH above its pl -    - Ve charge (Basic)

     

Protein extraction and Observation -

 

Extraction -       (in Buffer, solution).

Organ - tissue - cells - Homogenate - lysis with enzymes, Sonication -                  Centrifuge

 

Use techniques -

Spectrophotometer ( Absorb light at ~280nm )

( Absorbation directly proportional to the concentration of the solution - The Lambert-Beer Law )

  • Chromatography
  • Electrophoresis
  • Immunoassay

 

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