title: Use of hydrostatic pressure for modulation of protein chemical modification and enzymatic selectivity.
abstract: Using hydrostatic pressure to induce protein conformational changes can be a powerful tool for altering the availability of protein reactive sites and for changing the selectivity of enzymatic reactions. Using a pressure apparatus, it has been demonstrated that hydrostatic pressure can be used to modulate the reactivity of lysine residues of the protein ubiquitin with a water-soluble amine-specific homobifunctional coupling agent. Fewer reactive lysine residues were observed when the reaction was carried out under elevated pressure of 3 kbar, consistent with a pressure-induced conformational change of ubiquitin that results in fewer exposed lysine residues. Additionally, modulation of the stereoselectivity of an enzymatic transamination reaction was observed at elevated hydrostatic pressure. In one case, the minor diasteromeric product formed at atmospheric pressure became the major product at elevated pressure. Such pressure-induced alterations of protein reactivity may provide an important new tool for enzymatic reactions and the chemical modification of proteins.
authors.affiliation: Merck Sharp & Dohme, Merck Research Laboratories, Department of Process & Analytical Chemistry, Rahway, New Jersey 07065, USA. alexey.makarov@merck.com leo.joyce@merck.com.
authors.forename: Alexey A
authors.initials: AA
authors.lastname: Makarov
authors.affiliation: Merck Sharp & Dohme, Merck Research Laboratories, Department of Process & Analytical Chemistry, Rahway, New Jersey 07065, USA. alexey.makarov@merck.com leo.joyce@merck.com.
authors.forename: Roy
authors.initials: R
authors.lastname: Helmy
authors.affiliation: Merck Sharp & Dohme, Merck Research Laboratories, Department of Process & Analytical Chemistry, Rahway, New Jersey 07065, USA. alexey.makarov@merck.com leo.joyce@merck.com.
authors.forename: Leo
authors.initials: L
authors.lastname: Joyce
authors.affiliation: Merck Sharp & Dohme, Merck Research Laboratories, Department of Process & Analytical Chemistry, Rahway, New Jersey 07065, USA. alexey.makarov@merck.com leo.joyce@merck.com.
authors.forename: Mikhail
authors.initials: M
authors.lastname: Reibarkh
authors.affiliation: Merck Sharp & Dohme, Merck Research Laboratories, Department of Process & Analytical Chemistry, Rahway, New Jersey 07065, USA. alexey.makarov@merck.com leo.joyce@merck.com.
authors.forename: Mathew
authors.initials: M
authors.lastname: Maust
authors.affiliation: Merck Sharp & Dohme, Merck Research Laboratories, Department of Process & Analytical Chemistry, Rahway, New Jersey 07065, USA. alexey.makarov@merck.com leo.joyce@merck.com.
authors.forename: Sumei
authors.initials: S
authors.lastname: Ren
authors.affiliation: Merck Sharp & Dohme, Merck Research Laboratories, Department of Process & Analytical Chemistry, Rahway, New Jersey 07065, USA. alexey.makarov@merck.com leo.joyce@merck.com.
authors.forename: Ingrid
authors.initials: I
authors.lastname: Mergelsberg
authors.affiliation: Merck Sharp & Dohme, Merck Research Laboratories, Department of Process & Analytical Chemistry, Rahway, New Jersey 07065, USA. alexey.makarov@merck.com leo.joyce@merck.com.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Welch
mesh_terms: D000588:Amines / Q000737:chemistry; D000818:Animals; D004798:Enzymes / Q000737:chemistry* / Q000378:metabolism*; D006874:Hydrostatic Pressure; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism*; D012995:Solubility; D013379:Substrate Specificity; D014867:Water / Q000737:chemistry
chemical_list: D000588:Amines; D004798:Enzymes; D011506:Proteins; D014867:Water