title: End-label fingerprintings show that the N- and C-termini of actin are in the contact site with gelsolin.
abstract: Gelsolin was cleaved by chymotrypsin or thermolysin into an N-terminal Mr 45,000 fragment (45N) and a C-terminal Mr 38,000 fragment (38C). The N-terminal half was further cleaved into two fragments with Mr 17,000 (17N) and Mr 28,000 (28N). These fragments were complexed with actin and cross-linked with 1-ethyl-3-[3-(dimethylamino)prophyl]carbodiimide (EDC) to introduce covalent bonds into their contact sites. The location of these bonds was mapped along the actin sequence by end-label fingerprinting with highly sensitive probes for the N- and C-termini of actin. The mapping studies revealed that two gelsolin N-terminal fragments (17N and 28N) were cross-linked with the actin C-terminal segment. The result indicates that the actin N- and C-terminal segments are in the binding site of gelsolin.
authors.affiliation: Department of Biophysics and Biochemistry, Faculty of Science, University of Tokyo, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Sutoh
authors.forename: H L
authors.initials: HL
authors.lastname: Yin
mesh_terms: D000199:Actins / Q000032:analysis*; D001665:Binding Sites; D002135:Calcium-Binding Proteins / Q000032:analysis* / Q000502:physiology; D002918:Chymotrypsin; D003432:Cross-Linking Reagents; D003488:Cyanogen Bromide; D005022:Ethyldimethylaminopropyl Carbodiimide / Q000031:analogs & derivatives; D018260:Gelsolin; D006801:Humans; D006868:Hydrolysis; D015151:Immunoblotting; D008840:Microfilament Proteins / Q000032:analysis* / Q000502:physiology; D010446:Peptide Fragments / Q000032:analysis; D010449:Peptide Mapping; D012697:Serine Endopeptidases; D013820:Thermolysin
chemical_list: D000199:Actins; D002135:Calcium-Binding Proteins; D003432:Cross-Linking Reagents; D018260:Gelsolin; D008840:Microfilament Proteins; D010446:Peptide Fragments; C041829:1-ethyl-3-(3-(dimethylamino)propyl)carbodiimide methiodide; D012697:Serine Endopeptidases; D002918:Chymotrypsin; C036813:lysyl endopeptidase; D013820:Thermolysin; D003488:Cyanogen Bromide; D005022:Ethyldimethylaminopropyl Carbodiimide