Globin and linker sequences of the giant extracellular hemoglobin from the leech Macrobdella decora

• Published in: Journal of Protein Chemistry Vol. 22; no. 3; pp. 231 - 242
• Main Authors: Suzuki, Tomohiko, Vinogradov, Serge N
• Format: Journal Article
• Language: English
• Published: United States Springer Nature B.V 01.04.2003
• Subjects: Amino Acid Sequence; Animals; Aquatic life; Base Sequence; Cellular biology; Divergence; DNA, Complementary - genetics; Evolution; Evolution, Molecular; Freshwater; Gene duplication; Genetics; Globins - chemistry; Globins - genetics; Hemoglobin; Ionization; Lamellibrachia; Leeches; Leeches - chemistry; Leeches - genetics; Lumbricus; Macrobdella; Macrobdella decora; Mass spectrometry; Molecular Sequence Data; Mutation rates; Phylogeny; Sequence Alignment; Spectrometry
• ISSN: 0277-8033; 1572-3887; 1573-4943
• DOI: 10.1023/a:1025064318790

A detailed electrospray ionization mass spectrometric study of the approximately 3.5-MDa hexagonal bilayer hemoglobin (HBL Hb) from the pond leech Macrobdella decora has shown it to consist of at least six approximately 17-kDa globin chains, of which two are monomeric and the remaining four occur as disulfide-bonded heterodimers, and three approximately 24-kDa nonglobin linker chains (Weber et al., J. Mol. Biol. 251: 703-720, 1995). The cDNA sequences of the five major constituent chains, globin chains IIA, IIB, B, and C and linker chain L1, are reported here. The globins and linkers share 30%-50% and 20%-30% identity, respectively, with other annelid sequences. Furthermore, IIB and C align with strain A of annelid sequences, whereas IIA and B align with the strain B sequences. Although chains B and C are monomeric, chains IIA and IIB form the main disulfide-bonded dimer. They also have some unusual features: the distal His (E7) is replaced by Phe in IIA, and the highly conserved CD1Phe is replaced by Leu in IIB. In spite of these unusual features, the functional properties of Macrobdella Hb are comparable to those of other HBL Hbs. A phylogenetic analysis of the globin sequences from Macrobdella, the polychaete Tylorrhynchus, the oligochaete Lumbricus, and the vestimentiferan Lamellibrachia, indicates that the two strains originated by gene duplication followed by additional duplication of each of the two strains. The mutation rate of the linkers appeared to be faster than that of the globin chains. The phylogenetic trees constructed using the Maximum Likelihood, Neighbor-Joining and Fitch methods showed the Macrobdella globin sequences to be closest to Lumbricus, in agreement with a view of annelid evolution in which the divergence of the polychaetes occurred before the divergence of the leeches from oligochaetes.