Tyrosine 27 of the specificity polypeptide of EcoKI can be UV crosslinked to a bromodeoxyuridine-substituted DNA target sequence
| Autor: | CHEN, A, POWELL, L M, DRYDEN, D T F, MURRAY, N E, BROWN, T, Dryden, David |
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| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
Ultraviolet Rays
Protein subunit Molecular Sequence Data Biology In Vitro Techniques Substrate Specificity chemistry.chemical_compound Genetics Trypsin Amino Acid Sequence Tyrosine Binding site Peptide sequence Sequence (medicine) Binding Sites Base Sequence Sequence Homology Amino Acid technology industry and agriculture DNA DNA Restriction Enzymes Molecular biology Thymine Restriction enzyme Cross-Linking Reagents Biochemistry chemistry Bromodeoxyuridine |
| Zdroj: | CHEN, A, POWELL, L M, DRYDEN, D T F, MURRAY, N E, BROWN, T & Dryden, D 1995, ' TYROSINE-27 OF THE SPECIFICITY POLYPEPTIDE OF ECOKI CAN BE UV CROSS-LINKED TO A BROMODEOXYURIDINE-SUBSTITUTED DNA TARGET SEQUENCE ', Nucleic Acids Research, vol. 23, no. 7, pp. 1177-1183 . https://doi.org/10.1093/nar/23.7.1177 |
| DOI: | 10.1093/nar/23.7.1177 |
| Popis: | The specificity (S) subunit of the restriction enzyme EcoKI imparts specificity for the sequence AAC(N6)GTGC. Substitution of thymine with bromodeoxyuridine in a 25 bp DNA duplex containing this sequence stimulated UV light-induced covalent crosslinking to the S subunit. Crosslinking occurred only at the residue complementary to the first adenine in the AAC sequence, demonstrating a close contact between the major groove at this sequence and the S subunit. Peptide sequencing of a proteolytically-digested, crosslinked complex identified tyrosine 27 in the S subunit as the site of crosslinking. This is consistent with the role of the N-terminal domain of the S subunit in recognizing the AAC sequence. Tyrosine 27 is conserved in the S subunits of the three type I enzymes that share the sequence AA in the trinucleotide component of their target sequence. This suggests that tyrosine 27 may make a similar DNA contact in these other enzymes. |
| Databáze: | OpenAIRE |
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