Head-head Interactions Research Articles

Cyclic Continuous Glycation Enhanced Dispersibility of Myofibrillar Protein: Reaction Efficiency and Sites Modification.

Reaction efficiency in glycation lacks sufficient attention, leading to the waste of process costs. Cyclic continuous glycation (CCG) is an effective approach to accelerate covalent binding between myofibrillar protein (MP) and glucose. This study elucidated that CCG promoted the exposure of reactive glycated sites in MP with full unfolding of secondary and tertiary structures. Notably, the glycation rate was significantly increased by 65.43%. Physicochemical properties indicated that MP-glucose conjugates with high graft degree exhibited favorable solubility, dispersibility, and thermal stability. Furthermore, proteomics was applied to reveal the glycated sites and products in glycoconjugates of MP. Glycation preferentially acted on the tails of the myosin heavy chain. The glucosylation modification on the head region was enhanced by CCG contributing to the inhibition of the head-head interaction. Overall, this study systematically clarifies the mechanism of CCG, providing a theoretical basis for the application of glycation in innovative meat products.

Enhancing magnetically driven microswimmer velocity via low Reynolds number hydrodynamic interactions

The motion of comoving magnetic microswimmers is modeled by considering the inter-hydrodynamic interactions (HI) under low Reynolds number conditions. The microswimmer is a two-link design consisting of a magnetic head attached to a slender tail via a torsional spring, and it is driven by an external planar oscillatory magnetic field. The inter-HI considered are the head-head and tail-tail interactions. The propulsion velocity for the comoving mode is calculated and compared with that of an isolated mode. The comparative results show that the comoving mode velocity can be either similar or greater than the isolated mode, depending on the actuation frequency. The parametric dependency results show that the velocity generated in comoving mode depends on the average separation distance and length-to-width ratio of the tail. For proof of concept, a low-cost fabrication protocol is implemented to design a millimeter-sized magnetic flagellated swimmer. The experimental result shows that the comoving swimming mode generates larger velocity in comparison to isolated swimming.

Cryo-EM structure of tarantula myosin filament: Implications for analysis of myopathy mutations.

The high-resolution structure of myosin thick filaments of invertebrates has aided understanding of key muscle functions and analysis of disease mutations in humans. We previously published a low-resolution tarantula skeletal thick filament cryo-EM structure (Woodhead et al., 2005), which was used to map hypertrophic and dilated cardiomyopathy mutations (Alamo et al., 2017, Nag et al., 2017), present mostly in the head-head and head-tail interaction sites of the interacting-heads motif (IHM). This study suggested that the myosin coiled-coil tails in the filament backbone were arranged in groups of three, forming 40-Å-diameter subfilaments (Wray, 1979), but it lacked the detail required to resolve individual tails and to map the 40% of myopathy mutations that occur in the tail region of the molecule. Here we report a 5.5 Å resolution cryo-EM map of the tarantula thick filament backbone that resolves the densities of the full length of the myosin tails and their packing interactions. The map reveals that the tails do not form subfilaments but are arranged individually in layers forming a “molecular crystal,” in which the myosin coiled-coils gradually move towards the center of the filament, near the paramyosin core, while slowly curving around the filament axis. A similar arrangement was found in insect flight muscle thick filaments (Hu et al., 2016), suggesting it could be a general packing scheme in thick filaments, as originally proposed by Squire (1973). The positions of skip residues predict longer pitch regions and bending of the coiled-coils, as seen in the cryo-EM density. The structure also suggests new interactions of the blocked-head converter region with the backbone of the filament, implying hyperstability of IHMs. Improved resolution, and solution of the vertebrate thick filament structure, should enable precise mapping of tail mutations.

Simulation of surfactant adsorption at liquid-liquid interface: What we may expect from soft-core models?

The present work attempts to systematically explore the surfactant sorption at liquid-liquid interfaces with coarse-grained models targeting thermodynamic properties of reference liquid solutions. We employ dissipative particle dynamics with soft-core forcefield tested against experimental data on micellization of surfactants in water, and the previous results are reproduced in this work. We consider three different nonionic surfactants: hexaethylene glycol monododecyl ether (C12E6), 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol) known as Triton X-100 (TX-100), and two alkyl glucoside surfactants (CnG1) with n-alkane tail fragments and a saccharide hydrophilic head at decane-water and toluene-water interfaces. For TX-100, we composed a model based on the literature forcefield and found good agreement with the experimental critical micelle concentrations (CMCs). The head-head interactions are of different origins for different surfactant groups: entropic repulsion between ethylene oxide chains of C12E6 and TX-100, and more chemically specific and complex interactions between the maltose heads of alkyl glucosides. We interpret our results with the Redlich-Peterson equation of monolayer adsorption in order to relate the adsorption to the bulk concentration of the surfactant and the interfacial tension. The densities of the adsorbed monolayer at CMC mostly agree with the experimental data, and a reasonable agreement was obtained for the interfacial tension at CMC. At the same time, we found significant discrepancies between the simulated and experimental adsorption isotherms. We explain them by the oversimplified forcefield: when the parameters are fitted to the free energies of bulk solutions, they may not correctly reproduce the interfacial free energies.

All-atom molecular dynamics simulations reveal how kinesin transits from one-head-bound to two-heads-bound state.

Kinesin dimer walks processively along a microtubule (MT) protofilament in a hand-over-hand manner, transiting alternately between one-head-bound (1HB) and two-heads-bound (2HB) states. In 1HB state, one head bound by adenosine diphosphate (ADP) is detached from MT and the other head is bound to MT. Here, using all-atom molecular dynamics simulations we determined the position and orientation of the detached ADP-head relative to the MT-bound head in 1HB state. We showed that in 1HB state when the MT-bound head is in ADP or nucleotide-free state, with its neck linker being undocked, the detached ADP-head and the MT-bound head have the high binding energy, and after adenosine triphosphate (ATP) binds to the MT-bound head, with its neck linker being docked, the binding energy between the two heads is reduced greatly. These results reveal how the kinesin dimer retains 1HB state before ATP binding and how the dimer transits from 1HB to 2HB state after ATP binding. Key residues involved in the head-head interaction in 1HB state were identified.

Impact of Hypertrophic Cardiomyopathy Mutations on the Cardiac Myosin Super-Relaxed State

We aim to examine the effects of hypertrophic cardiomyopathy (HCM) associated mutations on the myosin super-relaxed state (SRX) through functional ATPase assays and time-resolved FRET structural measurements. Cardiac myosin isomerizes between the relaxed state (RX), which is available for activation and subsequent force generation with actin, and the kinetically inactive SRX, in which myosin is hypothesized to be structurally occluded from the force-generation cycle by the interacting-heads motif (IHM). We hypothesize that myosin regulatory light chain (RLC) mutations associated with HCM, specifically E22K and R58Q, cause hypercontractility through disruption of the myosin SRX. Based on proposed models of the IHM, these residues form stabilizing salt-bridge interactions with the opposite myosin. We hypothesize that mutations in the RLC associated with HCM increase myosin's basal ATPase activity due to disruption of SRX. To observe the effect of these mutations, we have performed ATPase assays to measure changes in myosin function, and time-resolved FRET to directly detect the changes in head-head interactions, due to changes in the population of SRX myosin. These measurements were performed on bovine cardiac myosin containing human cardiac RLC. This work was supported by NIH grants AR032961 and AR057220 (to DDT), and an American Heart Association Scientist Development Grant (to JMM).

CryoEM Single Particle Reconstruction of Dephosphorylated HMM from Smooth Muscle

The asymmetric head-head interaction that characterizes the inhibited state of smooth muscle myosin was first discovered in 3-D images of 2-D arrays formed on lipid monolayers at ∼2 nm resolution. The conformation is generally referred to as the “interacting heads motif” or IHM and has since been observed in 3-D images of thick filaments from all species examined from vertebrate striated muscle to the flight muscles of the large waterbug Lethocerus sp. indicating it is a ubiquitous myosin II inhibited conformation. However, current 3-D images reported from filaments are limited to resolutions lower than 1 nm. Here we report on progress toward an atomic resolution structure of the IHM of smooth muscle HMM using single particle methods from dispersions of single particles bound to a lipid monolayer. Data were collected on a Titan Krios equipped with a Volta Phase Plate and a Direct Electron DE-64 detector. Initial reconstructions, starting from a total of 1464 phase plate images from which 588,000 particles were selected using Gautomatch, which is a template matching particle picker, using a template derived from the smooth muscle HMM crystal structure. This structure went to 1.49 nm resolution from a subset of 285300 particles. The free and blocked heads of the IHM are easily distinguishable but the S2 domain is not yet visible. The structure is being refined with addition of new images. Supported by NIH.

Effects of different NaCl concentrations on self-assembly of silver carp myosin

The effect of NaCl (0.1 − 3.0 M) on self-assembly of silver carp myosin at 4 °C was investigated in terms of microstructure, conformation, intermolecular interactions, and particle size distribution. During setting at 4 °C, the self-assembly of myosin showed an obvious concentration dependence. At low concentrations (<0.3 M), myosin assembled into filaments mainly through rod-rod ionic linkages, and showed significantly (P < 0.05) higher turbidity and lower solubility than at ≥0.3 M. The addition of NaCl (0.3 − 0.6 M) led to the extension and dissolution of myosin by interfering with electrostatic interactions. The myosin assemblies gradually became smaller and more uniform. The solubility and Ca2+-ATPase activity reached a maximum at 0.6 M NaCl. When the concentrations increased to >1.0 M, myosin further assembled into filaments dominated by hydrophobic interactions. Both rod-rod and head-head interactions contributed to the myosin filaments at high concentrations (1.0 − 3.0 M). The solubility and uniformity of the myosin assemblies decreased as the NaCl concentration increased from 1.0 to 3.0 M.

Interacting-heads motif has been conserved as a mechanism of myosin II inhibition since before the origin of animals

Electron microscope studies have shown that the switched-off state of myosin II in muscle involves intramolecular interaction between the two heads of myosin and between one head and the tail. The interaction, seen in both myosin filaments and isolated molecules, inhibits activity by blocking actin-binding and ATPase sites on myosin. This interacting-heads motif is highly conserved, occurring in invertebrates and vertebrates, in striated, smooth, and nonmuscle myosin IIs, and in myosins regulated by both Ca2+ binding and regulatory light-chain phosphorylation. Our goal was to determine how early this motif arose by studying the structure of inhibited myosin II molecules from primitive animals and from earlier, unicellular species that predate animals. Myosin II from Cnidaria (sea anemones, jellyfish), the most primitive animals with muscles, and Porifera (sponges), the most primitive of all animals (lacking muscle tissue) showed the same interacting-heads structure as myosins from higher animals, confirming the early origin of the motif. The social amoeba Dictyostelium discoideum showed a similar, but modified, version of the motif, while the amoeba Acanthamoeba castellanii and fission yeast (Schizosaccharomyces pombe) showed no head-head interaction, consistent with the different sequences and regulatory mechanisms of these myosins compared with animal myosin IIs. Our results suggest that head-head/head-tail interactions have been conserved, with slight modifications, as a mechanism for regulating myosin II activity from the emergence of the first animals and before. The early origins of these interactions highlight their importance in generating the inhibited (relaxed) state of myosin in muscle and nonmuscle cells.

Myosin II Head Interaction in Primitive Species

Myosin II is a two-headed motor protein with an elongated α-helical tail. The motor domain of each head interacts with actin to convert the chemical energy of ATP into movement. Myosin II activity in muscle and nonmuscle cells is switched off by intramolecular interaction between its two heads, which inhibits their activity. This has been shown by EM and image processing of myosin filaments and isolated myosin molecules. In switched-off single molecules, the myosin tail folds into three segments, with the interacting heads folded back on the tail. The interacting-heads motif is highly conserved, being found in vertebrate and invertebrate smooth and striated muscle and in nonmuscle cells. We are investigating its evolutionary origins by EM imaging of isolated myosin molecules in the off-state. In previous work we found that the motif was present as far back as Cnidaria (sea anemones), the earliest animals with muscles. Here we have studied additional animal and non-animal species. At high (0.5 M) salt, all the myosin IIs showed the typical appearance of an extended tail and non-interacting heads. At low salt (0.15 M), under relaxing conditions (MgATP), insect indirect flight and embryonic skeletal muscle myosins showed a folded tail and similar head-head interactions to other muscles. Three species of primitive, non-animal myosins gave differing results. Acanthamoeba (reported previously) and Schizosaccharomyces pombe showed extended tails and no head-head interactions. The tails of these two myosins were approximately 30-40% shorter than the animal myosins, possibly accounting for their inability to fold. In contrast, Dictyostelium, with a tail 10% longer than animal myosin, showed head-head interactions similar to animal myosin; however, the conformation of the folded tail was different. These results suggest that head-head interaction arose before the evolution of animals.

The Inhibited, Interacting-Heads Motif Characterizes Myosin II from the Earliest Animals with Muscles

Electron microscopic studies have shown that the inhibited (relaxed) state of myosin II is characterized by an intramolecular interaction between myosin heads, in both thick filaments and isolated monomers. Interaction inhibits head activity by blocking actin-binding in one head and ATPase activity in the other. Interacting-heads are present in vertebrate smooth muscle and nonmuscle monomers, and in thick filaments of invertebrate smooth muscle (flatworms), invertebrate striated muscle (mollusks, arthropods) and vertebrate striated muscle (zebrafish, mouse and human); furthermore, head interactions underlie the off-state in myosins regulated by both Ca2+ binding and RLC phosphorylation. Head-head interaction has thus been conserved through evolution from mollusks and arthropods to humans. Our goal has been to determine how early this self-inhibiting motif arose, by studying the off-state of myosin monomers and/or filaments from primitive species. Filaments from sea anemones (Cnidaria), the most primitive animals with muscles, have the same 14.5-nm repeat as other muscles, but EM images have not yet been reconstructed. However, sea anemone myosin monomers, under relaxing conditions, have the same folded tail and bent-back, interacting-heads motif as myosins from the higher species, further demonstrating the very early origin of this structure. Interestingly, unlike the animals mentioned above, Acanthamoeba myosin monomers under relaxing conditions appear to lack a folded tail or interacting heads. The sequence of this myosin is strikingly different from the myosin IIs of animals, and its regulatory mechanism is quite different, suggesting that self-inhibition by interacting heads evolved after amoebae and animals diverged. Conservation of interacting heads over time suggests that it is a highly successful mechanism for regulating myosin activity. We are now studying the more primitive sea sponges (Porifera), one of the earliest animal groups, which lack muscles.

Role of the Essential Light Chain in the Activation of Smooth Muscle Myosin by Regulatory Light Chain Phosphorylation

The activity of smooth and non-muscle myosin II is regulated by phosphorylation of the regulatory light chain (RLC) at ser19. The dephosphorylated state of full-length monomeric myosin is characterized by an asymmetric intramolecular head-head interaction that completely inhibits the ATPase activity, accompanied by a hairpin fold of the tail, which prevents filament assembly. The mechanism by which ser19 phosphorylation disrupts these head-head interactions is unknown. Computational modeling (Tama et al., 2005. J. Mol. Biol. 345, 837-854) suggested that formation of the inhibited state is characterized by both torsional and bending motions about the myosin heavy chain (HC) at a location between the RLC and the essential light chain (ELC). Based on this model, placement of the regulatory domain at this locus might alter relative motions between the ELC and the RLC and thereby disrupt the inhibited state. Here we derive an atomic model based on this hypothesis for the phosphorylated state of the smooth muscle myosin light chain domain (LCD). We use a homology model for the structure of the RLC and a largely α-helical structure for the regulatory domain. This model predicts a set of specific interactions between the regulatory domain and both the myosin HC and the ELC. Site directed mutagenesis combined with ATPase and motility assays was used to show that interactions between the highly basic N-terminus of the RLC with acidic residues on helix-A of the ELC are required for phosphorylation to activate smooth muscle myosin. These sites are well conserved in all the myosin II ELC sequences examined despite the lack of known interacting partners and when substituted, are usually substituted with aspartic acid. Supported by NIH grants AR47421, AR53975 and HL38113 and RR12255.

Atomic-Level Visualization of Smooth Muscle Activation by Phosphorylation of the Myosin Regulatory Light Chain

We have engineered site-directed probe pairs in the regulatory light chain (RLC) for exchange into smooth muscle heavy meromyosin (HMM) and subfragment-1 (S1), in order to examine the phosphorylation-induced changes in RLC structural states using time-resolved FRET (TR-FRET). Phosphorylation of the RLC is required for activation of contraction in smooth muscle and modulates force in striated muscle. Force development in smooth muscle is triggered by phosphorylation of the RLC's N-terminal phosphorylation domain (PD) at Ser19, alleviating inhibitory interactions between the two heads. Since crystal structures of the RLC lack the PD, the mechanism by which RLC phosphorylation allosterically triggers disruption of HMM auto-inhibition remains unresolved. Our site-directed FRET probe pairs resolved intramolecular atomic distance measurements between the RLC core and the dynamic PD. Smooth muscle RLC assumes two structural states in both states of activation: phosphorylation shifts the equilibrium of these two structural states from primarily occupying the compact closed state to favoring the extended open state of the PD, relative to the RLC core. Compared to single-headed S1 (Kast et al., PNAS., 2010), the open state RLC conformation of unphosphorylated HMM is more compact, presumably due to head-head or RLC-RLC interactions. All-atom MD simulations corroborate our TR-FRET studies, and reveal specific salt-bridges that stabilize each structural state. The hypotheses generated from our MD simulations are being tested experimentally by TR-FRET via site-directed mutagenesis to manipulate charge. These studies offer the first atomic-resolution insight into the structural dynamics of RLC phosphorylation, and a similar approach should be applicable to striated muscle. Spectroscopy was performed in the Biophysical Spectroscopy Center at the University of Minnesota. This work was funded by NIH grants to DDT (R01 AR32961, P30 AR0507220, T32 AR007612).

Characterization of Three Full-length Human Nonmuscle Myosin II Paralogs

Nonmuscle myosin IIs (NM IIs) are a group of molecular motors involved in a wide variety of cellular processes including cytokinesis, migration, and control of cell morphology. There are three paralogs of the NM II heavy chain in humans (IIA, IIB, and IIC), each encoded by a separate gene. These paralogs are expressed at different levels according to cell type and have different roles and intracellular distributions in vivo. Most previous studies on NM II used tissue-purified protein or expressed fragments of the molecule, which presents potential drawbacks for characterizing individual paralogs of the intact protein in vitro. To circumvent current limitations and approach their native properties, we have successfully expressed and purified the three full-length human NM II proteins with their light chains, using the baculovirus/Sf9 system. The enzymatic and structural properties of the three paralogs were characterized. Although each NM II is capable of forming bipolar filaments, those formed by IIC tend to contain fewer constituent molecules than those of IIA and IIB. All paralogs adopt the compact conformation in the presence of ATP. Phosphorylation of the regulatory light chain leads to assembly into filaments, which bind to actin in the presence of ATP. The nature of interactions with actin filaments is shown with different paralogs exhibiting different actin binding behaviors under equivalent conditions. The data show that although NM IIA and IIB form filaments with similar properties, NM IIC forms filaments that are less well suited to roles such as tension maintenance within the cell.

Correction: Head-Head Interactions of Resting Myosin Crossbridges in Intact Frog Skeletal Muscles, Revealed by Synchrotron X-Ray Fiber Diffraction

Correction: Head-Head Interactions of Resting Myosin Crossbridges in Intact Frog Skeletal Muscles, Revealed by Synchrotron X-Ray Fiber Diffraction

Specific Charged Residues on the Myosin Heavy Chain (K368 and R406) Contribute to Head Interaction of Relaxed Smooth Muscle Myosin

Vertebrate smooth muscle contraction is regulated by phosphorylation of the myosin regulatory light chains. EM studies of smooth muscle myosin molecules suggest that in the dephosphorylated state, activity is switched off by asymmetric interactions between the two myosin heads, in which the “blocked” head is thought to be unable to bind to actin, while the “free” head has its ATPase activity inhibited. Comparative sequence analysis and molecular modeling suggest that the head interaction may be stabilized by charge attraction between D748 in the converter domain of the free head and K368 of the blocked head, and between R406 or K416 of the blocked head and E169 of the free head (Jung et al., MBC, 2008). To test these possibilities, we investigated the structure of expressed smooth muscle myosin and heavy meromyosin in which the charges at some of these sites was neutralized or reversed. While most of the mutants showed no obvious change in structure compared with wild type, HMM with charge reversal at both 368 and 406 showed fewer molecules with interacting heads when observed by metal shadowing. This was supported by a decrease in sedimentation coefficient (determined by analytical ultracentrifugation) and an increase in actin-activated ATPase activity of the mutant. Whole myosin with the same mutations formed filaments more readily than wild type, also consistent with these results. These observations suggest that charge interactions involving K368 and R406 play a role in stabilizing the head-head interaction of the inactive state. Interestingly, mutation of R403 in cardiac myosin (R406 in smooth muscle) increases motor activity of myosin and causes familial hypertrophic cardiomyopathy. Our results suggest that this could be in part a result of changes in head interaction.

Hydrogen-Deuterium Exchange Mass Spectrometry of Phosphorylation Induced Changes in Smooth Muscle Myosin

Inter-molecular interactions between two smooth muscle myosin heads have been observed using chemical cross linking and visualized by electron microscopy. In order to probe the nature of the phosphorylation-dependent head-head interactions, we performed Hydrogen-Deuterium exchange electrospray ionization mass spectrometry (HDX-MS) on phosphorylated and unphosphorylated smooth muscle heavy mero myosin (HMM) from chicken gizzard. HDX rates are determined by solvent accesibility and the strength of hydrogen bonding and thus are a good fingerprint of intra- and intermolecular structural changes. The observed decrease in deuterium uptake upon phosphorylation in the coiled-coil and the hinge region of the heavy chain (residues 864-877), is consistent with the unraveling of the myosin dimer at the hinge. Unexpectedly, we have observed phosphorylation dependent changes in the regulatory light chain (RLC 58-69), which are not implicated in the RLC-RLC or RLC-heavy chain interface. No changes were observed for N-terminal region of RLC that was postulated to undergo major secondary structure changes, or in the heavy chain actin binding site (residues 397-437), or any of the residues that form the head-head interface in the EM model of Wendt & Taylor. This latter finding indicates that the interaction between the heads is probably transient in solution, or in the unlikely case, interacting heads constitute a very small fraction of total protein.

放射光X線回折によるミオシンクロスブリッジの頭部間相互作用—ミオシンフィラメントの筋収縮制御の構造的基盤

放射光X線回折によるミオシンクロスブリッジの頭部間相互作用—ミオシンフィラメントの筋収縮制御の構造的基盤

Head-head interactions of resting myosin crossbridges in intact frog skeletal muscles, revealed by synchrotron x-ray fiber diffraction.

The intensities of the myosin-based layer lines in the x-ray diffraction patterns from live resting frog skeletal muscles with full thick-thin filament overlap from which partial lattice sampling effects had been removed were analyzed to elucidate the configurations of myosin crossbridges around the thick filament backbone to nanometer resolution. The repeat of myosin binding protein C (C-protein) molecules on the thick filaments was determined to be 45.33 nm, slightly longer than that of myosin crossbridges. With the inclusion of structural information for C-proteins and a pre-powerstroke head shape, modeling in terms of a mixed population of regular and perturbed regions of myosin crown repeats along the filament revealed that the myosin filament had azimuthal perturbations of crossbridges in addition to axial perturbations in the perturbed region, producing pseudo-six-fold rotational symmetry in the structure projected down the filament axis. Myosin crossbridges had a different organization about the filament axis in each of the regular and perturbed regions. In the regular region that lacks C-proteins, there were inter-molecular interactions between the myosin heads in axially adjacent crown levels. In the perturbed region that contains C-proteins, in addition to inter-molecular interactions between the myosin heads in the closest adjacent crown levels, there were also intra-molecular interactions between the paired heads on the same crown level. Common features of the interactions in both regions were interactions between a portion of the 50-kDa-domain and part of the converter domain of the myosin heads, similar to those found in the phosphorylation-regulated invertebrate myosin. These interactions are primarily electrostatic and the converter domain is responsible for the head-head interactions. Thus multiple head-head interactions of myosin crossbridges also characterize the switched-off state and have an important role in the regulation or other functions of myosin in thin filament-regulated muscles as well as in the thick filament-regulated muscles.

Role of metal ions in growth and stability of Langmuir-Blodgett films on homogeneous and heterogeneous surfaces

Structure and stability of cadmium arachidate (CdA) Langmuir-Blodgett (LB) films on homogeneous (i.e., OH-, H-passivated Si(001) substrates) and heterogeneous (i.e., Br-passivated Si(001) substrates) surfaces were studied using X-ray reflectivity and atomic force microscopy techniques and compared with those of nickel arachidate (NiA) LB films. While on OH-passivated Si, an asymmetric monolayer (AML) structure starts to grow, on H-passivated Si, a symmetric monolayer (SML) of CdA forms, although for both the films, pinhole-type defects are present as usual. However, on heterogeneous Br-passivated Si substrates, a combination of AML, SML, shifted SML and SML on top of AML (i.e., AML/SML), all types of structures are found to grow in such a way that, due to the variation of heights in the out-of-plane direction, ring-shaped in-plane nanopatterns of CdA molecules are generated. Probably due to stronger head-head interactions and higher metal ion-carboxylic ligand bond strength for CdA molecules compared to NiA, easy flipping of SML on top of another preformed SML, i.e. a SML/SML structure formation was not possible and as a result a wave-like modulation is observed for the CdA film on such heterogeneous substrate. The presence of hydrophilic/hydrophobic interfacial stress on the heterogeneous substrate thus modifies the deposited molecular structure so that the top surface morphology for a CdA film is similar to monolayer buckling while that for NiA film is similar to monolayer collapse.