Isolation and characterization of mosquito ferritin and cloning of a cDNA that encodes one subunit.

B. C. Dunkov, D. Zhang, K. Choumarov, Joy Winzerling, J. H. Law

Research output: Contribution to journalArticle

63 Citations (Scopus)

Abstract

Ferritin, an iron storage protein, was isolated from larvae and pupae of Aedes aegypti grown in an iron-rich medium. Mosquito ferritin is a high molecular weight protein composed of several different, relatively small, subunits. Subunits of molecular mass 24, 26, and 28 kDa are equally abundant, while that of 30 kDa is present only in small amounts. The N-terminal sequence of the 24 and 26 kDa subunits are identical for the first 30 amino acids, while that of the 28 kDa subunit differs. Studies using antiserum raised against a subunit mixture showed that the ferritin subunit were present in larvae, pupae, and adult females, and were increased in animals exposed to excess iron. The antiserum also was used to screen a cDNA library from unfed adult female mosquitoes. Nine clones were obtained that differed only in a 27 bp insertion in the 3' end. Rapid amplification of cDNA ends (RACE) was used to obtain the complete protein coding sequence. A putative iron-responsive element (IRE) is present in the 5'-untranslated region. The deduced amino acid sequence shows a typical leader sequence, consistent with the fact that most insect ferritins are secreted, rather than cytoplasmic proteins. The sequence encodes a mature polypeptide of 20,566 molecular weight, smaller than the estimated size of any of the subunits. However, the sequence exactly matches the N-terminal sequences of the 24 and 26 kDa subunits as determined by Edman degradation. Of the known ferritin sequences, that of the mosquito is most similar to that of somatic cells of a snail.

Original languageEnglish (US)
Pages (from-to)293-307
Number of pages15
JournalArchives of Insect Biochemistry and Physiology
Volume29
Issue number3
StatePublished - 1995

Fingerprint

Cloning
ferritin
Ferritins
Culicidae
Organism Cloning
molecular cloning
Complementary DNA
Iron
iron
Pupa
molecular weight
Larva
antiserum
pupae
Immune Sera
Proteins
Molecular Weight
Molecular weight
Amino Acids
rapid amplification of cDNA ends

ASJC Scopus subject areas

  • Insect Science
  • Biochemistry, Genetics and Molecular Biology(all)
  • Physiology
  • Physiology (medical)

Cite this

Isolation and characterization of mosquito ferritin and cloning of a cDNA that encodes one subunit. / Dunkov, B. C.; Zhang, D.; Choumarov, K.; Winzerling, Joy; Law, J. H.

In: Archives of Insect Biochemistry and Physiology, Vol. 29, No. 3, 1995, p. 293-307.

Research output: Contribution to journalArticle

@article{c8ff75db0f4746a6953a712c75c95cf8,
title = "Isolation and characterization of mosquito ferritin and cloning of a cDNA that encodes one subunit.",
abstract = "Ferritin, an iron storage protein, was isolated from larvae and pupae of Aedes aegypti grown in an iron-rich medium. Mosquito ferritin is a high molecular weight protein composed of several different, relatively small, subunits. Subunits of molecular mass 24, 26, and 28 kDa are equally abundant, while that of 30 kDa is present only in small amounts. The N-terminal sequence of the 24 and 26 kDa subunits are identical for the first 30 amino acids, while that of the 28 kDa subunit differs. Studies using antiserum raised against a subunit mixture showed that the ferritin subunit were present in larvae, pupae, and adult females, and were increased in animals exposed to excess iron. The antiserum also was used to screen a cDNA library from unfed adult female mosquitoes. Nine clones were obtained that differed only in a 27 bp insertion in the 3' end. Rapid amplification of cDNA ends (RACE) was used to obtain the complete protein coding sequence. A putative iron-responsive element (IRE) is present in the 5'-untranslated region. The deduced amino acid sequence shows a typical leader sequence, consistent with the fact that most insect ferritins are secreted, rather than cytoplasmic proteins. The sequence encodes a mature polypeptide of 20,566 molecular weight, smaller than the estimated size of any of the subunits. However, the sequence exactly matches the N-terminal sequences of the 24 and 26 kDa subunits as determined by Edman degradation. Of the known ferritin sequences, that of the mosquito is most similar to that of somatic cells of a snail.",
author = "Dunkov, {B. C.} and D. Zhang and K. Choumarov and Joy Winzerling and Law, {J. H.}",
year = "1995",
language = "English (US)",
volume = "29",
pages = "293--307",
journal = "Archives of Insect Biochemistry and Physiology",
issn = "0739-4462",
publisher = "John Wiley and Sons Ltd",
number = "3",

}

TY - JOUR

T1 - Isolation and characterization of mosquito ferritin and cloning of a cDNA that encodes one subunit.

AU - Dunkov, B. C.

AU - Zhang, D.

AU - Choumarov, K.

AU - Winzerling, Joy

AU - Law, J. H.

PY - 1995

Y1 - 1995

N2 - Ferritin, an iron storage protein, was isolated from larvae and pupae of Aedes aegypti grown in an iron-rich medium. Mosquito ferritin is a high molecular weight protein composed of several different, relatively small, subunits. Subunits of molecular mass 24, 26, and 28 kDa are equally abundant, while that of 30 kDa is present only in small amounts. The N-terminal sequence of the 24 and 26 kDa subunits are identical for the first 30 amino acids, while that of the 28 kDa subunit differs. Studies using antiserum raised against a subunit mixture showed that the ferritin subunit were present in larvae, pupae, and adult females, and were increased in animals exposed to excess iron. The antiserum also was used to screen a cDNA library from unfed adult female mosquitoes. Nine clones were obtained that differed only in a 27 bp insertion in the 3' end. Rapid amplification of cDNA ends (RACE) was used to obtain the complete protein coding sequence. A putative iron-responsive element (IRE) is present in the 5'-untranslated region. The deduced amino acid sequence shows a typical leader sequence, consistent with the fact that most insect ferritins are secreted, rather than cytoplasmic proteins. The sequence encodes a mature polypeptide of 20,566 molecular weight, smaller than the estimated size of any of the subunits. However, the sequence exactly matches the N-terminal sequences of the 24 and 26 kDa subunits as determined by Edman degradation. Of the known ferritin sequences, that of the mosquito is most similar to that of somatic cells of a snail.

AB - Ferritin, an iron storage protein, was isolated from larvae and pupae of Aedes aegypti grown in an iron-rich medium. Mosquito ferritin is a high molecular weight protein composed of several different, relatively small, subunits. Subunits of molecular mass 24, 26, and 28 kDa are equally abundant, while that of 30 kDa is present only in small amounts. The N-terminal sequence of the 24 and 26 kDa subunits are identical for the first 30 amino acids, while that of the 28 kDa subunit differs. Studies using antiserum raised against a subunit mixture showed that the ferritin subunit were present in larvae, pupae, and adult females, and were increased in animals exposed to excess iron. The antiserum also was used to screen a cDNA library from unfed adult female mosquitoes. Nine clones were obtained that differed only in a 27 bp insertion in the 3' end. Rapid amplification of cDNA ends (RACE) was used to obtain the complete protein coding sequence. A putative iron-responsive element (IRE) is present in the 5'-untranslated region. The deduced amino acid sequence shows a typical leader sequence, consistent with the fact that most insect ferritins are secreted, rather than cytoplasmic proteins. The sequence encodes a mature polypeptide of 20,566 molecular weight, smaller than the estimated size of any of the subunits. However, the sequence exactly matches the N-terminal sequences of the 24 and 26 kDa subunits as determined by Edman degradation. Of the known ferritin sequences, that of the mosquito is most similar to that of somatic cells of a snail.

UR - http://www.scopus.com/inward/record.url?scp=0029176319&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029176319&partnerID=8YFLogxK

M3 - Article

C2 - 7655055

AN - SCOPUS:0029176319

VL - 29

SP - 293

EP - 307

JO - Archives of Insect Biochemistry and Physiology

JF - Archives of Insect Biochemistry and Physiology

SN - 0739-4462

IS - 3

ER -