Binding is not enough: flexibility is needed for photocrosslinking of Lck kinase by benzophenone photoligands.
Benzophenone photophores are employed widely for photoaffinity-labeling studies. Photolabeling with benzophenone, however, is hardly a routine experiment. Even when a photoprobe binds to its target, photocrosslinking does not necessarily occur. This is because photolabeling by benzophenone is affected by many factors other than target-binding, such as conformational flexibility of photoligand. Despite the widespread recognition of such complications, there has been no systematic study to assess the relative importance of individual factors that can affect photolabeling efficiency. In order to gain an insight into this problem, we conducted a structure-activity relationship (SAR) study of benzophenone photoligands for Lck kinase, in which photoligands with varying target-binding affinity and conformational flexibility were compared. The study found that binding-affinity, as indicated by kinase inhibitory potency, did not correlate with photolabeling efficiency. Instead, conformational flexibility was found to be the determining factor for efficient photolabeling by our photoligands. Implication of the current findings, in particular, with regard to selection and optimization of benzophenone photoligands, is discussed.
Kawamura
Akira
A
Department of Chemistry, Hunter College of CUNY, 695 Park Avenue, New York, NY 10065, USA. akawamur@hunter.cuny.edu
Hindi
Sagit
S
Mihai
Doina M
DM
James
Laurence
L
Aminova
Olga
O
D000595:Amino Acid Sequence; D001577:Benzophenones / Q000138:chemical synthesis / Q000494:pharmacology*; D001665:Binding Sites; D015153:Blotting, Western; D004791:Enzyme Inhibitors / Q000138:chemical synthesis / Q000494:pharmacology*; D008024:Ligands; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck) / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D019920:Photoaffinity Labels / Q000138:chemical synthesis / Q000494:pharmacology*; D013329:Structure-Activity Relationship
D001577:Benzophenones; D004791:Enzyme Inhibitors; D008024:Ligands; D019920:Photoaffinity Labels; C047723:benzophenone; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck)
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