Chicken GRIFIN: A homodimeric member of the galectin network with canonical properties and a unique expression profile.
Occurrence of the adhesion/growth-regulatory galectins as family sets the challenge to achieve a complete network analysis. Along this route taken for a well-suited model organism (chicken), we fill the remaining gap to characterize its seventh member known from rat as galectin-related inter-fiber protein (GRIFIN) in the lens. Its single-copy gene is common to vertebrates, with one or more deviations from the so-called signature sequence for ligand (lactose) contact. The chicken protein is a homodimeric agglutinin with capacity to bind β-galactosides, especially the histo-blood group B tetrasaccharide, shown by solid-phase/cell assays and a glycan microarray. Mass spectrometric identification of two lactose-binding peptides after tryptic on-bead fragmentation suggests an interaction at the canonical region despite a sequence change from Arg to Val at the site, which impairs reactivity of human galectin-1. RT-PCR and Western blot analyses of specimen from adult chicken organs reveal restriction of expression to the lens, here immunohistochemically throughout its main body. This report sets the stage for detailed structure-activity studies to define factors relevant for affinity beyond the signature sequence and to perform the first complete network analysis of the galectin family in developing and adult organs of a vertebrate.
Institute of Physiological Chemistry, Faculty of Veterinary Medicine, Ludwig-Maximilians-University Munich, Veterinärstr. 13, 80539 Munich, Germany.
Gabriel
G
García Caballero
Institute of Physiological Chemistry, Faculty of Veterinary Medicine, Ludwig-Maximilians-University Munich, Veterinärstr. 13, 80539 Munich, Germany.
Herbert
H
Kaltner
Department of Applied Tumor Biology, Institute of Pathology, Medical School of the Ruprecht-Karls-University, Im Neuenheimer Feld 224, 69120 Heidelberg, Germany.
Malwina
M
Michalak
Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya, Moscow, Russia.
Nadezhda
N
Shilova
Pharma Biotech Development Penzberg, Roche Diagnostics GmbH, 82377 Penzberg, Germany.
Michelle
M
Yegres
Institute of Physiological Chemistry, Faculty of Veterinary Medicine, Ludwig-Maximilians-University Munich, Veterinärstr. 13, 80539 Munich, Germany.
Sabine
S
André
Institute of Physiological Chemistry, Faculty of Veterinary Medicine, Ludwig-Maximilians-University Munich, Veterinärstr. 13, 80539 Munich, Germany.
Anna-Kristin
AK
Ludwig
Institute of Physiological Chemistry, Faculty of Veterinary Medicine, Ludwig-Maximilians-University Munich, Veterinärstr. 13, 80539 Munich, Germany.
Joachim C
JC
Manning
Institute of Physiological Chemistry, Faculty of Veterinary Medicine, Ludwig-Maximilians-University Munich, Veterinärstr. 13, 80539 Munich, Germany.
Sebastian
S
Schmidt
Genomics and Proteomics Core Facility, German Cancer Research Center, 69120 Heidelberg, Germany.
Martina
M
Schnölzer
Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya, Moscow, Russia.
Nicolai V
NV
Bovin
Pharma Biotech Development Penzberg, Roche Diagnostics GmbH, 82377 Penzberg, Germany.
Dietmar
D
Reusch
Department of Applied Tumor Biology, Institute of Pathology, Medical School of the Ruprecht-Karls-University, Im Neuenheimer Feld 224, 69120 Heidelberg, Germany.
Jürgen
J
Kopitz
Institute of Physiological Chemistry, Faculty of Veterinary Medicine, Ludwig-Maximilians-University Munich, Veterinärstr. 13, 80539 Munich, Germany. Electronic address: gabius@tiph.vetmed.uni-muenchen.de.
Hans-Joachim
HJ
Gabius
D000595:Amino Acid Sequence; D000818:Animals; D030161:Avian Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D001665:Binding Sites / Q000235:genetics; D015153:Blotting, Western; D002645:Chickens; D005136:Eye Proteins / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D037161:Galectins / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D020869:Gene Expression Profiling / Q000379:methods*; D053263:Gene Regulatory Networks; D006801:Humans; D007150:Immunohistochemistry; D007785:Lactose / Q000378:metabolism; D007908:Lens, Crystalline / Q000378:metabolism; D010802:Phylogeny; D011485:Protein Binding; D055503:Protein Multimerization; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid
D030161:Avian Proteins; D005136:Eye Proteins; D037161:Galectins; D011994:Recombinant Proteins; D007785:Lactose
Fiber cells; In-source decay; Lectin; Lens; Phylogenesis