Phosphorylation of hydroxylysine residues in collagen synthesized by cultured aortic smooth muscle cells

Journal Article · Wed May 01 00:00:00 EDT 1985 · Proc. Natl. Acad. Sci. U.S.A.; (United States)

DOI:https://doi.org/10.1073/pnas.82.10.3091· OSTI ID:6335492

Urushizaki, Y; Seifter, S

OV-Phosphohydroxylysine was chemically synthesized and techniques were established for its identification by combined use of cation-exchange chromatography, thin-layer electrophoresis at pH 1.9 and 3.5, and thin-layer chromatography. Conditions were also determined to permit hydrolysis of proteins in 2 M HCl without loss of the phosphono group of phosphohydroxylysine residues. Experiments were then performed showing that TSP was incorporated into the hydroxylysine residues of cell-associated collagens when cultured calf aorta medial smooth muscle cells were incubated with (TSP)orthophosphate. In other experiments, the cells incorporated (TH)lysine into hydroxylysine residues of cell-associated collagen and then TSP into phosphohydroxylysine residues. The doubly labeled phosphohydroxylysine subsequently isolated showed nearly 1:1 stoichiometry with respect to incorporation of precursor lysine and phosphorus. Finally, in preliminary experiments done with a cell-free extract of the smooth muscle cells, TSP was transferred from (el-TSP)ATP to hydroxylysine residues in several kinds of collagenous substrates. Thus, this work shows that smooth muscle cells have the capacity to phosphorylate hydroxylysine residues in their cell-associated collagens and provides preliminary evidence that a protein kinase is involved.

• Research Organization: Yeshiva Univ., Bronx, NY
• OSTI ID: 6335492
• Journal Information: Proc. Natl. Acad. Sci. U.S.A.; (United States), Vol. 82:10
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
LYSINE
PHOSPHORYLATION
MUSCLES
BIOCHEMISTRY
ORGANIC PHOSPHORUS COMPOUNDS
CHEMICAL PREPARATION
PHOSPHORUS 32
TRACER TECHNIQUES
TRITIUM COMPOUNDS
AMINO ACIDS
ANIMAL CELLS
AORTA
COLLAGEN
DOUBLE LABELLING
ELECTROPHORESIS
IN VITRO
ION EXCHANGE CHROMATOGRAPHY
PHOSPHATES
THIN-LAYER CHROMATOGRAPHY
ARTERIES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BLOOD VESSELS
BODY
CARBOXYLIC ACIDS
CARDIOVASCULAR SYSTEM
CHEMICAL REACTIONS
CHEMISTRY
CHROMATOGRAPHY
DAYS LIVING RADIOISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
LABELLED COMPOUNDS
LABELLING
LIGHT NUCLEI
NUCLEI
ODD-ODD NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANS
OXYGEN COMPOUNDS
PHOSPHORUS COMPOUNDS
PHOSPHORUS ISOTOPES
PROTEINS
RADIOISOTOPES
SCLEROPROTEINS
SEPARATION PROCESSES
SYNTHESIS
550201* - Biochemistry- Tracer Techniques