protein engineering 🤡🤡🤡🤡🤡🤡

Question 1

antigen

Answer 1

Molecules that cause an immune response (recognized and bound by antibodies)

Question 2

antibodies

Answer 2

Proteins of the immune system that recognize and bind to foreign molecules

Question 3

phage display library

Answer 3

collection of bacteriophage particles that have segments of foreign proteins protruding from their surface

Question 4

combinatorial library

Answer 4

Large series of related molecules that have been systematically generated by combining chemical groups and/or molecular motifs

Question 5

alternative splicing library

Answer 5

Library of gene or protein sequences generated by randomized inclusion or exclusion of exons from an original protein

Question 6

combinatorial screening

Answer 6

Screening of a large series of related molecules for those with useful properties

Question 7

random shuffling library

Answer 7

Library of gene or protein sequences generated by randomized shuffling and linking of short segments

Question 8

How are protein libraries created?

Answer 8

DNA is chemically synthesized and then the modules are assembled to give new artificial genes with PCR methods (usually overlapping primers).

Question 9

DNA shuffling

Answer 9

Method of artificial evolution in which genes are cut into segments that are mutagenized, mixed, shuffled, and rejoined - trying to generate novel or improved protein activities.

Question 10

epitopes

Answer 10

Localized regions of an antigen to which an antibody binds

BODER DEF: peptide sequence for which a monoclonal antibody will bind to (a flag so antibodies can recognize)

Question 11

How does recombining protein domains affect proteins?

Answer 11

Proteins have discrete domains with individual functions. Recombining domains from different proteins can generate hybrid proteins with novel combinations of properties.

Question 12

Directed Evolution

Answer 12

Technique for enhancing or altering the original activity of an enzyme by randomly mutating the gene and then screening for the new activity - library construction

Question 13

error-prone PCR

Answer 13

Type of PCR in which mutations are introduced at random during the amplification steps - used for random mutagenesis

Question 14

tyrosyl-tRNA synthetase

Answer 14

e Enzyme that charges tRNA with tyrosine (mutated through site-specific directed evolution)

Question 15

How is the industrial efficiency of proteins increased?

Answer 15

Some proteins are larger than necessary so extra sequences are removed and the protein is simplified.

Question 16

How do manipulations to the protein active site alter the stability of the protein?

Answer 16

altering amino acids in and around the active site may change the specificity of an enzyme toward substrates or cofactors.

A cofactor is a non- protein chemical compound that is bound to a protein and is required for the protein’s biological activity.

Question 17

How can you increase protein stability with amino acids?

Answer 17

Replacing glycine with any other amino acid or increasing the number of proline residues in a polypeptide chain will increase stability.

Question 18

How do certain unstable amino acids affect protein stability?

Answer 18

High temperature or extreme pH can convert asparagine and glutamine amides to their corresponding acids. The replacement of a neutral amid by the negatively charged carboxyl negatively impacts structure and activity.

Question 19

disulfide bonds

Answer 19

A bond between two closely situated cysteine residues in a protein that stabilizes the tertiary structure

Question 20

Melting Temperature (Tm)

Answer 20

the temperature at which 50% of the protein is denatured