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Structural organization of the lens fiber cell plasma membrane protein MP18

Journal Article · · Journal of Biological Chemistry; (USA)
OSTI ID:5037932
; ; ; ; ; ;  [1]
  1. Univ. of Minnesota, St. Paul (USA)
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The 18,000-dalton bovine lens fiber cell intrinsic membrane protein MP18 was phosphorylated on a serine residue by both cAMP-dependent protein kinase and protein kinase C. In addition, this protein bound calmodulin and was recognized by a monoclonal antibody (2D10). These different regions were localized using enzymatic and chemical fragmentation of electrophoretically purified MP18 that had been phosphorylated with either cAMP-dependent protein kinase or protein kinase C. Partial digestion of 32P-labeled MP18 with protease V8 resulted in a Mr = 17,000 peptide that bound calmodulin, but neither contained 32P or was recognized by the monoclonal antibody 2D10. Furthermore, the 17-kDa peptide had the same N-terminal amino acid sequence as MP18. Thus, the monoclonal antibody 2D10 recognition site and the protein kinase phosphorylation site(s) are close together and confined to a small region in the C terminus of MP18. This conclusion was confirmed in experiments where MP18 was fragmented with trypsin, endoproteinase Lys-C, or CNBr. The location of the phosphorylation site was confirmed by sequencing the small 32P-labeled, C-terminal peptide that resulted from protease V8 digestion of 32P-labeled MP18. This peptide contained a consensus sequence for cAMP-dependent protein kinase.
OSTI ID:
5037932
Journal Information:
Journal of Biological Chemistry; (USA), Journal Name: Journal of Biological Chemistry; (USA) Vol. 264:33; ISSN JBCHA; ISSN 0021-9258
Country of Publication:
United States
Language:
English

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Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMINO ACID SEQUENCE
ANIMALS
AUTORADIOGRAPHY
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BODY
BODY AREAS
CELL CONSTITUENTS
CELL MEMBRANES
CHEMICAL REACTIONS
CRYSTALLINE LENS
DAYS LIVING RADIOISOTOPES
ELECTROPHORESIS
ENZYMES
EYES
FACE
HEAD
HYDROLASES
ISOTOPES
LIGHT NUCLEI
MAMMALS
MEMBRANE PROTEINS
MEMBRANES
MICE
MOLECULAR STRUCTURE
MOLECULAR WEIGHT
NUCLEI
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANS
PEPTIDE HYDROLASES
PEPTIDES
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PHOSPHORYLATION
PROTEINS
RADIOISOTOPES
RODENTS
SENSE ORGANS
SERINE PROTEINASES
VERTEBRATES