Analysis of the membrane-interacting domain of the sea urchin sperm adhesive protein bindin

Journal Article · Tue Nov 14 00:00:00 EST 1989 · Biochemistry; (USA)

DOI:https://doi.org/10.1021/bi00449a029· OSTI ID:7137901

Kennedy, L; DeAngelis, P L; Glabe, C G ^[1]

---

1. Univ. of California, Irvine (USA)

The authors have investigated the domain of the bindin polypeptide the selectively associates with gel-phase phospholipid vesicles. They found that small trypsin fragments of bindin retain the ability to selectively associate with gel-phase vesicles. The primary amino acid sequence of bindin suggests that these peptides are derived from the central portion of the polypeptide between residues 77 and 126, which is the most hydrophobic region of bindin. They have also employed 3-(trifluoromethyl)-3-(m-({sup 125}I)iodophenyl)diazirine (TID) and novel, radioiodinated, photoactivatable derivatives of the polar head group of phosphatidylethanolamine (ASD-PE and ASA-PE) to identify membrane-associated polypeptide segments after the transfer of radiolabel from the probe to the bindin polypeptide. After photolysis, bindin was selectively labeled only from probes incorporated in gel-phase vesicles. The labeling of bindin was much more efficient from the head group probes ASA-PE and ASD-PE (8 and 2% of the total label, respectively) in comparison to the hydrophobic probe TID (less than 0.02% of the total label), suggesting that bindin is localized within the polar part of the bilayer. Protease mapping experiments with V8 protease, trypsin, and endoprotease Lys-C suggest that some of the probe label is distributed along the amino-terminal portion of bindin between residues 1 and 76 and the rest of the label is restricted to the segments between residues 77 and 126 which also selectively bind to gel-phase vesicles. The carboxyl-terminal portion of bindin residues 127 and 236 is not labeled.

• OSTI ID: 7137901
• Journal Information: Biochemistry; (USA), Vol. 28:23; ISSN 0006-2960
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
CELL MEMBRANES
CROSS-LINKING
MEMBRANE PROTEINS
AMINO ACID SEQUENCE
PHOSPHOLIPIDS
DERIVATIZATION
IODINE 125
PEPTIDE HYDROLASES
POLYPEPTIDES
SEA URCHINS
SODIUM IODIDES
SPERMATOZOA
THIN-LAYER CHROMATOGRAPHY
TRYPSIN
ALKALI METAL COMPOUNDS
ANIMALS
AQUATIC ORGANISMS
BETA DECAY RADIOISOTOPES
CELL CONSTITUENTS
CHEMICAL REACTIONS
CHROMATOGRAPHY
DAYS LIVING RADIOISOTOPES
ECHINODERMS
ELECTRON CAPTURE RADIOISOTOPES
ENZYMES
ESTERS
GAMETES
GERM CELLS
HALIDES
HALOGEN COMPOUNDS
HYDROLASES
INORGANIC PHOSPHORS
INTERMEDIATE MASS NUCLEI
INVERTEBRATES
IODIDES
IODINE COMPOUNDS
IODINE ISOTOPES
ISOTOPES
LIPIDS
MEMBRANES
MOLECULAR STRUCTURE
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
ORGANIC PHOSPHORUS COMPOUNDS
PEPTIDES
PHOSPHORS
POLYMERIZATION
PROTEINS
RADIOISOTOPES
SEPARATION PROCESSES
SERINE PROTEINASES
SODIUM COMPOUNDS
550201* - Biochemistry- Tracer Techniques