BCM 251: Introduction to Proteins - Lecture 2 Study Notes

BCM 251: Introduction to Proteins - Lecture 2: Classification and Properties of Amino Acids

Professor Oleg Reva, Chapters 3.1


Structures and Properties of Amino Acids

  • All amino acids contain:

    • One alpha-amino group

    • One alpha-carboxyl group

    • Differ in the chemical properties of their side radicals (R-groups)

  • In proteins, amino acids are linked:

    • By a covalent bond known as a peptide bond

    • The term residue signifies the loss of a water molecule (dehydration reaction) during peptide bond formation

  • Average molecular weight (MW) of amino acid residues = 110 Da

    • MW of an individual amino acid residue = MW of the amino acid - 18 Da (to account for the water molecule)

  • Carbon atoms of R-residues are numbered from the carbon attached to the amino and carboxyl groups (a-carbon) using Greek letters (b, g, d, e) or numeric numbering from the carboxyl carbon atom.


Exercises

Exercise 1:

  • A protein with an MW of 30,900 Da has 2% cysteine residues by weight.

    • Cys MW = 121 Da

    • Question: How many cysteine residues does the protein contain?


Amino Acid Classification Based on R-groups

Group One: Nonpolar (Aliphatic) R-groups

  • Includes the following amino acids:

    • Glycine (Gly, G)

    • Alanine (Ala, A)

    • Leucine (Leu, L)

    • Isoleucine (Ile, I)

    • Valine (Val, V)

    • Proline (Pro, P)

    • Methionine (Met, M)

  • Key Points:

    • Aliphatic refers to the hydro-carbonic nature of the side chains lacking aromatic (benzene) rings.

    • Hydrophobicity increases with the number of carbon atoms in the hydrocarbon chain.

    • Generally found inside proteins, except alanine and glycine are ambivalent; they can be located both inside and on the surface of proteins.


Proline
  • Proline has an aliphatic circular R-group where the terminal carbon is attached to the a-nitrogen.

    • Ring structure is referred to as an imine group, making proline an imino acid.

    • Reduces structural flexibility and bands protein chains in proteins.

Methionine
  • Methionine contains a sulfur atom within its hydrocarbon side chain.

    • It is hydrophobic and found within the hydrophobic cores of proteins.

    • Although it has a sulfur atom, it remains chemically inactive with no specific roles beyond helping with protein lipid interactions and stabilizing hydrophobic regions in proteins.


Group Two: Aromatic R-groups

  • Includes:

    • Phenylalanine

    • Tyrosine

    • Tryptophan

  • Key Features:

    • Contain phenyl, hydroxyphenyl, and indole aromatic rings respectively.

    • Contribute to hydrophobic interactions, ordered by relative hydrophobicity: Phe > Trp > Tyr.

    • Tyrosine and tryptophan can form hydrogen bonds due to –OH and –NH- groups.

  • Important for specific protein interactions with DNA and organic compounds.

  • Function as precursors of various neurotransmitters.


UV Light Absorbance
  • Tryptophan and tyrosine absorb UV light, leading to characteristic strong absorbance at 280 nm.

    • This property is useful for detecting proteins in solution, particularly against contamination in DNA/RNA samples at 260 nm (260/280 ratio).


Group Three: Polar Uncharged R-groups

  • Includes:

    • Serine (Ser, S)

    • Threonine (Thr, T)

    • Cysteine (Cys, C)

    • Glutamine (Gln, Q)

    • Asparagine (Asn, N)

  • Features:

    • These amino acids are common on the surface of water-soluble proteins.

    • Serine and threonine have a polar hydroxyl (—OH) group which allows them to form hydrogen bonds with water, enhancing protein hydrophilicity.


Cysteine
  • Contains a thiol group (—SH)

    • Can react with another cysteine to form a disulfide bridge (—S—S—), also known as cystine, which is hydrophobic.

  • Plays a crucial role in stabilizing tertiary protein structures and participates in biochemical reduction reactions controlling cellular redox potential.


Glutamine and Asparagine
  • Contain amide groups derived from carboxyl groups of glutamic and aspartic acids.

    • Not ionizable within the pH range of cellular conditions, hence they have no charge.

  • Act as donors of amino groups in synthesizing various nitrogen-containing molecules.

  • Capable of forming hydrogen bonds in protein structures.

  • Can be hydrolyzed back to their precursor acids during post-translational modifications.


Group Four: Positively Charged R-groups

  • Includes:

    • Histidine (His, H)

    • Lysine (Lys, K)

    • Arginine (Arg, R)

  • Characterization:

    • These amino acids have alkaline side chains, positively charged at neutral pH.

    • Charges arise from an additional amino group in lysine; a guanidine group in arginine; and an imidazole ring in histidine.

  • Histidine has a pKa ≈ 6.0 making it capable of existing in both protonated and unprotonated states, affecting protein properties and facilitating enzyme activity.


Group Five: Negatively Charged R-groups

  • Includes:

    • Glutamic acid (Glu, E)

    • Aspartic acid (Asp, D)

  • Features:

    • Have extra carboxyl groups in their side chains in addition to the alpha-carboxyl.

    • At neutral pH, these groups deprotonate to form carboxylate anions (glutamate and aspartate), hence they have a negative charge.

  • Glu and Asp are strongly hydrophilic and participate in interactions with positively charged amino acids and ligands.


Common Proteinogenic Amino Acids

  • Proteinogenic amino acids:

    • Amino acids that are incorporated biosynthetically into protein chains by ribosomes.

    • A total of 20 common proteinogenic amino acids exist, encoded by specific codons in DNA/mRNA.

    • L-isomers are proteinogenic; D-isomers are not.

  • Rare or uncommon proteinogenic amino acids resulting from enzymatic modification occur post-translationally.


Uncommon/Rare Proteinogenic Amino Acids

  • Selenocysteine:

    • Present in all three domains of life involved in oxidative stress enzymes.

    • Abbreviated as Sec; encoded by the UGA stop codon plus a Sec insertion sequence in mRNA.

  • Selenomethionine:

    • Found in plants responsive to oxidative stress; can affect horses' health.

    • Abbreviated as Met; encoded by AUG (same as methionine).

  • Pyrrolysine:

    • Found in some methanogenic bacteria and archaea, abbreviated as Pyl; encoded by UAG stop codon and requires an additional enzyme for synthesis.


Non-Proteinogenic Amino Acids

  • Non-proteinogenic amino acids:

    • Cannot be found within proteins.

    • D-optical isomers of amino acids are typically non-proteinogenic, but common in bioactive peptides produced by bacteria, fungi, and plants, such as in peptidoglycan.

  • Notable examples:

    • b-Alanine: Precursor of vitamin B5 and coenzyme A, also naturally occurs in antioxidant peptides like carnosine and anserine.

    • Other non-proteinogenic amino acids include ornithine, citrulline, and S-adenosylmethionine (SAM).


Post-Translational Modifications of Amino Acids

  • Common modifications leading to amino acid derivatives in proteins:

    1. Hydroxyproline and Hydroxylysine: Found in collagen.

    2. Thyroxine: Present in thyroglobulin in thyroid glands.

    3. Phosphorylation: Occurs on Ser, Thr, and Tyr side chains, and on His, Lys, Arg, Asp, and Glu.

    4. Methylation and Acetylation: Seen in histones, contributing to epigenetic regulation of gene transcription.

    5. Glutamate Methylation: Part of bacterial chemotaxis, regulating bacterial movement by methylation of flagella motor enzymes.

    6. Carboxylation of Glutamate: Important in the coagulation cascade.

    7. Desmosine: Synthesized from 4 lysine residues, integral to elastin function in providing tissue elasticity.


Exercises and Questions for Self-Control

Exercise 2: Identify amino acid(s) with given characteristics:
a. Positively charged at neutral pH: Arg, Lys, His
b. Has alcohol or phenolic group: Tyr
c. Contains thiol: Cys
d. Aromatic character: Phe, Tyr, Trp
e. Contains amide group: Asn, Gln
f. Participants in hydrogen bonding: Multiple amino acids including Ser, Thr, Cys, etc.

Exercise 3: List amino acids with specific characteristics:
a. Positively charged at pH 7: Arg, Lys, His
b. Negatively charged at pH 7: Asp, Glu
c. Can form disulfide bonds: Cys
d. Aromatic character: Tyr, Phe, Trp
e. Not chiral: Gly
f. More than one chiral carbons: Thr, Ile, etc.

Questions for Self-Control:

  • Name at least 4 amino acids that do not occur in proteins: b-Alanine, Ornithine, Citrulline, SAM.


Conclusion

These study notes synthesize the classification and properties of amino acids essential in understanding protein structure and function. The detailed notes can serve as a comprehensive resource for further studies in protein biochemistry.