title: Spectroscopic properties of Carcinus aestuarii hemocyanin and its structural subunits.
abstract: Hemocyanin (Hc) of Carcinus aestuarii contains three major and one minor electrophoretically separable polypeptide chains which were purified by fast protein liquid chromatography (FPLC) ion exchange chromatography. N-terminal amino acid sequences of four structural subunits (SSs) from C. aestuarii were compared with known N-terminal sequences from other arthropodan hemocyanins. The conformational changes, induced by various treatments, were monitored by far UV, CD and fluorescence spectroscopy. The critical temperatures for the structural subunits, Tc, determined by fluorescence spectroscopy, are in the region of 52-59 degrees C and coincide with the melting temperatures, Tm (49-55 degrees C), determined by CD spectroscopy. The free energy of stabilization in water, delta GDH2O, toward guanidinium hydrochloride is about 1.3 times higher for the dodecameric Hc as compared to the isolated subunits and about one time higher for Cal, comparing with other SSs. The studies reveal that the conformational stability of the native dodecamer towards various denaturants (temperature and guanidinium hydrochloride) indicate that the quaternary structure is stabilized by oligomerization between structural subunits, and the possibility of a structural role of the sugar mojeties cannot be excluded.
authors.affiliation: Institute of Organic Chemistry, Bulgarian Academy of Sciences, Sofia, Bulgaria. pda54@hotmail.com
authors.forename: P
authors.initials: P
authors.lastname: Dolashka-Angelova
authors.forename: R
authors.initials: R
authors.lastname: Hristova
authors.forename: S
authors.initials: S
authors.lastname: Stoeva
authors.forename: W
authors.initials: W
authors.lastname: Voelter
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001181:Arthropods / Q000737:chemistry*; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D006433:Hemocyanins / Q000737:chemistry*; D008969:Molecular Sequence Data; D010100:Oxygen / Q000737:chemistry; D011487:Protein Conformation; D016415:Sequence Alignment; D013050:Spectrometry, Fluorescence; D013696:Temperature; D013816:Thermodynamics
chemical_list: D006433:Hemocyanins; D010100:Oxygen