Source of error in the chromatographic study of /sup 35/S-sulfate labeled mucous glycoproteins secreted by the gill epithelium of Mytilus edulis

Journal Article · Wed Jan 01 04:00:00 EST 1986 · Prep. Biochem.; (United States)

OSTI ID:5073309

Sabouni, A H; Ma, J K; Malanga, C J

HPLC combined with (/sup 35/S)-sulfate/(/sup 3/H)-glucosamine radiolabeling were employed to study the synthesis and secretion of mucous glycoproteins. The secreted radiolabeled glycoproteins were separated from the medium by precipitation with a mixture of trichloroacetic-phosphotungstic acids (TCA/PTA). The redissolved glycoproteins were chromatographed on an anion exchange protein column at varying pH of the mobile phase and fractions were collected for liquid scintillation counting. Varying the pH of the mobile phase from pH 3 to 7 resulted in a decrease of glycoprotein bound (/sup 35/S) from 69.5 to 0.5% of the total recovered (/sup 35/S)-sulfate with the remainder recovered as free (/sup 35/S)-sulfate. The (/sup 3/H)-labeled glycoprotein recovered under the uV peaks at this pH range was 99.5%. When high performance size exclusion chromatography was performed the change in mobile phase pH did not affect the 100% recovery of either (/sup 35/S)-or (/sup 3/H)-labels under the uV peaks. No free (/sup 35/S)-sulfate was obtained when (/sup 35/S)-labeled glycoproteins were separated from the medium using dialysis. These data suggest that the standard method of TCA/PTA precipitation of (/sup 35/S)-labeled glycoproteins may cleave the (/sup 35/S)-sulfate ester linkages to the oligosaccharide chains. The (/sup 35/S)-sulfate may then rebind to the macromolecule by a relatively strong noncovalent bond. This may prove critical in anion exchange protein HPLC studies.

• Research Organization: WVU Medical Center, Morgantown
• OSTI ID: 5073309
• Journal Information: Prep. Biochem.; (United States), Journal Name: Prep. Biochem.; (United States) Vol. 3; ISSN PRBCB
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANIMAL TISSUES
ANIMALS
AQUATIC ORGANISMS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BODY
CARBOHYDRATES
CHROMATOGRAPHY
COUNTING TECHNIQUES
DAYS LIVING RADIOISOTOPES
EPITHELIUM
ERRORS
EVEN-ODD NUCLEI
GILLS
GLUCOPROTEINS
INVERTEBRATES
ISOTOPE APPLICATIONS
ISOTOPES
LABELLED COMPOUNDS
LIGHT NUCLEI
MOLLUSCS
MUSSELS
NUCLEI
ORGANIC COMPOUNDS
PROTEINS
RADIOISOTOPES
RESPIRATORY SYSTEM
SACCHARIDES
SCINTILLATION COUNTING
SECRETION
SEPARATION PROCESSES
SULFUR 35
SULFUR ISOTOPES
TISSUES
TRACER TECHNIQUES