Conversion of proteins to diazeniumdiolate-based nitric oxide donors.
Michael reaction of the methoxymethyl-protected monodiazeniumdiolate of piperazine (MOM-PIPERAZI/NO) with 4-maleimidobutyric acid followed by its conversion to the N-hydroxy-succinimido ester produces a reagent capable of transferring the nitric oxide (NO)-donating diazeniumdiolate group to the terminal amines of the lysine residues contained in proteins. The reagent has been used to produce diazeniumdiolated bovine serum albumin (D-BSA) and diazeniumdiolated human serum albumin (D-HSA) containing 22 and 19 modified lysyl groups, respectively. Upon dissolution in pH 7.4 phosphate buffer at 37 degrees C, these albumin derivatives gradually released all of their contained NO (approximately 40 mol/mol of protein) with initial rates of about 30-40 pmol/min/mg and half-lives on the order of 3 weeks. This methodology is now available for use in exploiting the unique specific metabolic interactions of proteins to target NO therapy to specific physiological processes in vivo.
Hrabie
J A
JA
Chemical Synthesis and Analysis Laboratory and Intramural Research Support Program, SAIC Frederick, NCI-FCRDC, Frederick, Maryland 21702, USA. hrabie@mail.ncifcrf.gov
Saavedra
J E
JE
Roller
P P
PP
Southan
G J
GJ
Keefer
L K
LK
D003432:Cross-Linking Reagents / Q000737:chemistry; D006801:Humans; D006834:Hydrazines / Q000737:chemistry*; D013058:Mass Spectrometry; D009569:Nitric Oxide / Q000737:chemistry; D020030:Nitric Oxide Donors / Q000138:chemical synthesis* / Q000737:chemistry; D010879:Piperazines / Q000737:chemistry; D012709:Serum Albumin / Q000737:chemistry*; D012710:Serum Albumin, Bovine / Q000737:chemistry; D013388:Succinimides / Q000737:chemistry
D003432:Cross-Linking Reagents; D006834:Hydrazines; D020030:Nitric Oxide Donors; D010879:Piperazines; D012709:Serum Albumin; D013388:Succinimides; D012710:Serum Albumin, Bovine; D009569:Nitric Oxide; C032138:N-(gamma-maleimidobutyryloxy)succinimide