Identification by integrated computer modeling and light scattering studies of an electrostatic serum albumin-hyaluronic acid binding site.
Dynamic light scattering and turbidimetry, carried out on solutions of hyaluronic acid (HA) and bovine or human serum albumin (SA) at fixed ionic strength (I), revealed a critical pH corresponding to the onset of HA-SA soluble complex formation. Subsequent reduction of pH below pH(c), corresponding to an increase in protein net positive charge, results in phase separation of the complex. The sensitivity of pH(c) to I indicated the primacy of electrostatic interactions in this process. Since pH(c) was always above the pK(a) of HA, these effects could be attributed to the influence of protein charge. The electrostatic potential around HSA was modeled using DelPhi (MSI) under pH, I conditions corresponding to incipient binding, phase separation, and noninteraction. At all incipient binding conditions (i.e., pH(c), at varying I), an identical region of positive potential 5 A from the protein van der Waals surface appeared. This unique domain intensified with a decrease in pH or I (corresponding to stronger binding), and diminished with an increase in pH or I (i.e., at noninteracting conditions). The size and low curvature of this domain could readily accommodate a 12 nm (decamer) sequence of HA. Simple electrostatic considerations indicate an electrostatic binding energy for the formation of this complex of ca. 1 kT, consistent with the condition of incipient complex formation. We suggest that such weak electrostatic binding may characterize nonspecific interactions for other protein-gylcosaminoglycan pairs.
Grymonpré
K R
KR
Department of Chemistry, Indiana University-Purdue University, Indianapolis, Indiana 46202-3276, USA.
Staggemeier
B A
BA
Dubin
P L
PL
Mattison
K W
KW
D000818:Animals; D001665:Binding Sites; D002417:Cattle; D003198:Computer Simulation; D006801:Humans; D006820:Hyaluronic Acid / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D008027:Light; D008958:Models, Molecular; D012542:Scattering, Radiation; D012709:Serum Albumin / Q000737:chemistry*; D012995:Solubility; D055672:Static Electricity
D012709:Serum Albumin; D006820:Hyaluronic Acid