End-label fingerprintings show that the N- and C-termini of actin are in the contact site with gelsolin

Kazuo Sutoh, Helen L. Yin

Research output: Contribution to journalArticle

25 Citations (Scopus)

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)propyl]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 N-terminal segment, while the gelsolin C-terminal fragment (38C) was 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.

Original languageEnglish (US)
Pages (from-to)5269-5275
Number of pages7
JournalBiochemistry
Volume28
Issue number12
StatePublished - 1989

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Gelsolin
Labels
Actins
Thermolysin
Covalent bonds
Chymotrypsin
Binding Sites

ASJC Scopus subject areas

  • Biochemistry

Cite this

End-label fingerprintings show that the N- and C-termini of actin are in the contact site with gelsolin. / Sutoh, Kazuo; Yin, Helen L.

In: Biochemistry, Vol. 28, No. 12, 1989, p. 5269-5275.

Research output: Contribution to journalArticle

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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)propyl]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 N-terminal segment, while the gelsolin C-terminal fragment (38C) was 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.",
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N2 - 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)propyl]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 N-terminal segment, while the gelsolin C-terminal fragment (38C) was 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.

AB - 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)propyl]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 N-terminal segment, while the gelsolin C-terminal fragment (38C) was 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.

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