Collagen-binding growth factors: Production and characterization of functional fusion proteins having a collagen-binding domain

Nozomu Nishi, Osamu Matsushita, Kouichi Yuube, Hiroshi Miyanaka, Akinobu Okabe, Fumio Wada

Research output: Contribution to journalArticle

84 Citations (Scopus)

Abstract

The autocrine/paracrine peptide signaling molecules such as growth factors have many promising biologic activities for clinical applications. However, one cannot expect specific therapeutic effects of the factors administered by ordinary drug delivery systems as they have limited target specificity and short half-lives in vivo. To overcome the difficulties in using growth factors as therapeutic agents, we have produced fusion proteins consisting of growth factor moieties and a collagen-binding domain (CBD) derived from Clostridium histolyticum collagenase. The fusion proteins carrying the epidermal growth factor (EGF) or basic fibroblast growth factor (bFGF) at the N terminal of CBD (CBEGF/CBFGF) tightly bound to insoluble collagen and stimulated the growth of BALB/c 3T3 fibroblasts as much as the unfused counterparts. CBEGF, when injected subcutaneously into nude mice, remained at the sites of injection for up to 10 days, whereas EGF was not detectable 24 h after injection. Although CBEGF did not exert a growth- promoting effect in vivo, CBFGF, but not bFGF, strongly stimulated the DNA synthesis in stromal cells at 5 days and 7 days after injection. These results indicate that CBD may be used as an anchoring unit to produce fusion proteins nondiffusible and long-lasting in vivo.

Original languageEnglish
Pages (from-to)7018-7023
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume95
Issue number12
DOIs
Publication statusPublished - Jun 9 1998
Externally publishedYes

Fingerprint

Intercellular Signaling Peptides and Proteins
Collagen
Fibroblast Growth Factor 2
Epidermal Growth Factor
Injections
Microbial Collagenase
Proteins
Paracrine Communication
Therapeutic Uses
Drug Delivery Systems
Stromal Cells
Growth
Nude Mice
Fibroblasts
Peptides
DNA
Therapeutics

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Collagen-binding growth factors : Production and characterization of functional fusion proteins having a collagen-binding domain. / Nishi, Nozomu; Matsushita, Osamu; Yuube, Kouichi; Miyanaka, Hiroshi; Okabe, Akinobu; Wada, Fumio.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 95, No. 12, 09.06.1998, p. 7018-7023.

Research output: Contribution to journalArticle

@article{8574d33fbefb4bdea2104eebff157ba7,
title = "Collagen-binding growth factors: Production and characterization of functional fusion proteins having a collagen-binding domain",
abstract = "The autocrine/paracrine peptide signaling molecules such as growth factors have many promising biologic activities for clinical applications. However, one cannot expect specific therapeutic effects of the factors administered by ordinary drug delivery systems as they have limited target specificity and short half-lives in vivo. To overcome the difficulties in using growth factors as therapeutic agents, we have produced fusion proteins consisting of growth factor moieties and a collagen-binding domain (CBD) derived from Clostridium histolyticum collagenase. The fusion proteins carrying the epidermal growth factor (EGF) or basic fibroblast growth factor (bFGF) at the N terminal of CBD (CBEGF/CBFGF) tightly bound to insoluble collagen and stimulated the growth of BALB/c 3T3 fibroblasts as much as the unfused counterparts. CBEGF, when injected subcutaneously into nude mice, remained at the sites of injection for up to 10 days, whereas EGF was not detectable 24 h after injection. Although CBEGF did not exert a growth- promoting effect in vivo, CBFGF, but not bFGF, strongly stimulated the DNA synthesis in stromal cells at 5 days and 7 days after injection. These results indicate that CBD may be used as an anchoring unit to produce fusion proteins nondiffusible and long-lasting in vivo.",
author = "Nozomu Nishi and Osamu Matsushita and Kouichi Yuube and Hiroshi Miyanaka and Akinobu Okabe and Fumio Wada",
year = "1998",
month = "6",
day = "9",
doi = "10.1073/pnas.95.12.7018",
language = "English",
volume = "95",
pages = "7018--7023",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "12",

}

TY - JOUR

T1 - Collagen-binding growth factors

T2 - Production and characterization of functional fusion proteins having a collagen-binding domain

AU - Nishi, Nozomu

AU - Matsushita, Osamu

AU - Yuube, Kouichi

AU - Miyanaka, Hiroshi

AU - Okabe, Akinobu

AU - Wada, Fumio

PY - 1998/6/9

Y1 - 1998/6/9

N2 - The autocrine/paracrine peptide signaling molecules such as growth factors have many promising biologic activities for clinical applications. However, one cannot expect specific therapeutic effects of the factors administered by ordinary drug delivery systems as they have limited target specificity and short half-lives in vivo. To overcome the difficulties in using growth factors as therapeutic agents, we have produced fusion proteins consisting of growth factor moieties and a collagen-binding domain (CBD) derived from Clostridium histolyticum collagenase. The fusion proteins carrying the epidermal growth factor (EGF) or basic fibroblast growth factor (bFGF) at the N terminal of CBD (CBEGF/CBFGF) tightly bound to insoluble collagen and stimulated the growth of BALB/c 3T3 fibroblasts as much as the unfused counterparts. CBEGF, when injected subcutaneously into nude mice, remained at the sites of injection for up to 10 days, whereas EGF was not detectable 24 h after injection. Although CBEGF did not exert a growth- promoting effect in vivo, CBFGF, but not bFGF, strongly stimulated the DNA synthesis in stromal cells at 5 days and 7 days after injection. These results indicate that CBD may be used as an anchoring unit to produce fusion proteins nondiffusible and long-lasting in vivo.

AB - The autocrine/paracrine peptide signaling molecules such as growth factors have many promising biologic activities for clinical applications. However, one cannot expect specific therapeutic effects of the factors administered by ordinary drug delivery systems as they have limited target specificity and short half-lives in vivo. To overcome the difficulties in using growth factors as therapeutic agents, we have produced fusion proteins consisting of growth factor moieties and a collagen-binding domain (CBD) derived from Clostridium histolyticum collagenase. The fusion proteins carrying the epidermal growth factor (EGF) or basic fibroblast growth factor (bFGF) at the N terminal of CBD (CBEGF/CBFGF) tightly bound to insoluble collagen and stimulated the growth of BALB/c 3T3 fibroblasts as much as the unfused counterparts. CBEGF, when injected subcutaneously into nude mice, remained at the sites of injection for up to 10 days, whereas EGF was not detectable 24 h after injection. Although CBEGF did not exert a growth- promoting effect in vivo, CBFGF, but not bFGF, strongly stimulated the DNA synthesis in stromal cells at 5 days and 7 days after injection. These results indicate that CBD may be used as an anchoring unit to produce fusion proteins nondiffusible and long-lasting in vivo.

UR - http://www.scopus.com/inward/record.url?scp=0032499796&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0032499796&partnerID=8YFLogxK

U2 - 10.1073/pnas.95.12.7018

DO - 10.1073/pnas.95.12.7018

M3 - Article

C2 - 9618531

AN - SCOPUS:0032499796

VL - 95

SP - 7018

EP - 7023

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 12

ER -